An automated heat treatment controller consistently controls heat treating a metallic target in a furnace according to a heat treatment recipe including a cycle temperature and a cycle exposure time. The automated heat treatment controller includes an operator interface for prompting an operator to load or remove a heat treated target. The controller further provides for commanding furnace door locks and for monitoring system performance. An automated heat treatment furnace method includes the steps of pre-heating the furnace, prompting the user to load the material, reheating the furnace to heat treatment parameters, controlling the exposure time and temperature, and prompting the removal of the material.

Grieve ovens, also referred to as furnaces, are utilized to process materials at elevated temperatures for homogenization, recrystallization, die pressing, annealing, stress relief, and hot rolling. These heat treatment processes provide methods of controlling grain structures, grain sizes, distributions of alloying elements, and grain orientations. Desirable sizes and shapes are obtained by rolling after the heat treatments. Manual operations are employed to control all heat treatment processes. Operators set the cycle temperature at the setpoint, or the "aim temperature," of the temperature controller 30. When the oven is heated to a temperature close to the cycle temperature, materials are loaded in the oven for heat treatment. Heat treatment time begins to count once the oven door is closed.
This manual process has many disadvantages, which leads to improperly heat treated materials and thus waste or defective products.
Exposure time to the cycle temperature might not be consistently controlled. This can be due to the operator failing to initiate the timer properly or to remove the material promptly when cycle time has expired. Since recipe parameters are manually referenced, the operator may reference the cycle time incorrectly.
Variations in temperature control occur since process control is subject to the discretion of individual operator. Mistakes happen for many reasons, such as when the operator incorrectly references the appropriate heat treatment recipe. A different operator assumes control of an ongoing treatment at shift change and may make a mistake as to the temperature parameters. The operator may take more or less than time than average in loading the material, allowing the starting temperature to vary from the expected temperature due to the door being open. Undetected degraded hardware could cause loss of control over the cycle temperature.
This lack of real-time monitoring creates additional disadvantages of the manual system. The lack of an automatic alert wastes time causing an economic loss. Relying on an individual operator to monitor the process closely causes increased labor expense in staffing and training. Moreover, the mundane nature of such manual monitoring makes timely detection difficult.
Improvements using statistical process control are difficult to accomplish since historical temperature data is not automatically gathered. As is the real-time data, the historical data is subject to the errors during manual collection. Moreover, errors are introduced when inputting the manually-collected data for analyzing for statistical process control (SPC) purposes. These additional steps make the time delay and frequency in such calculations problematic.
Therefore, a significant need continues to exist for a manner of controlling heat treatment furnaces to reduce amount of improperly processed materials and to reduce the labor expense.


A method of searching for and obtaining a vaccine against the pathogenic effects related to the infection of an animal or human host by a retrovirus that penetrates into a target cell of the host, and a vaccine obtained by the method are provided. The method includes preparing candidate vaccine agents based on a polypeptide comprising at least part of an envelope protein of a pathogenic strain of the retrovirus and selecting as the vaccine a modified polypeptide chosen from polypeptides that induces an immune response directed against an immunodominant region of an envelope protein of the retrovirus and not against a protein of the host.

The present invention relates to a method for obtaining vaccines for preventing the pathogenic effects related, in humans and in vertebrate animals, to retroviral infections.
The pathogenic effects related to a retroviral infection are the harmful effects, including possible oncogenic or immunosuppressive effects, induced by the introduction of a retrovirus into the body of a host (mammal, bird or alternatively fish), followed by the penetration and by the replication of said retrovirus in the cells of the host which are target cells for the retrovirus, that is to say cells into which the virus is capable of penetrating.
Retroviruses are thus named because they have the capacity, by virtue of the enzyme called reverse transcriptase, of carrying out transcription of RNA to DNA, whereas in living beings, the genetic information usually goes from the DNA of the chromosomes to proteins, via messenger RNA.
Three subfamilies can be distinguished in the retroviral family: the oncoviruses, the lentiviruses and the spumaviruses.
The oncoviruses are retroviruses thus termed because they can be associated with cancers and malignant infections. There may be mentioned, for example, leukemogenic viruses (such as the avian leukemia virus (ALV), the murine leukemia virus (MULV), also called Moloney virus, the feline leukemia virus (FELV), human leukemia viruses such as HTLV1 and HTLV2, the simian leukemia virus or STLV, the bovine leukemia virus or BLV), the primate type D oncoviruses, the type B oncoviruses which are inducers of mammary tumors, or oncoviruses which cause a rapid cancer (such as the Rous sarcoma virus or RSV); see for example STEHELIN et al., J. Mol. Biol. 101: 349-365 (1976).
The lentiviruses are thus named because they are responsible for slow-progressing pathological conditions which very frequently involve immunosuppressive phenomena, including AIDS.
The appended Table 1 indicates, by way of illustration, the pathological conditions associated with some lentiviruses, as well as the main target cells for these lentiviruses.
The spumaviruses manifest fairly low specificity for a given cell type or a given species, and they are sometimes associated with immunosuppressive phenomena; that is the case, for example, for the simian foamy virus (or SFV).
One of the aims of the present invention is the development of methods and vaccine products intended for effectively preventing the pathogenic effects, including the oncogenic or immunosuppressive effects, related to the infection of a host organism by a retrovirus.
Immunosuppression related to infection has been observed for a large number of retroviruses, and may be considered as a pathogenic constant of retroviral infection; see in particular BENDINELLI et al., Advances in Cancer Research 45: 125-181 (1985). This is the case in particular for lentivirus infections. It is also the case in a good number of oncovirus infections; see for example P. SONIGO in the book xe2x80x9cSIDA et infection par VIHxe2x80x9d [AIDS and HIV Infection], MONTAGNIER et al. (Mxc3xa9decine Science Flammarion), pages 113-122 (1989).
Many human and animal vaccines have been tested for preventing the pathogenic effects of retrovirus infections but, as a general rule, these vaccines are not very effective or are ineffective. In particular, in the field of human or animal AIDS, it is observed that, 14 years after the discovery of the HIV virus (BARRE-SINOUSSI et al., Science 220: 868-871, 1983), it has not yet been possible to find a vaccine which is able to effectively stop a post-vaccine HIV or SIV infection; see for example LINHART et al., AIDS Research and Human Retroviruses 13: 593-599 (1997); VOGT et al., Vaccine 13: 202-208 (1995); and LETVIN et al., J. Virol. 69: 4569-4571 (1995).
The majority of the vaccine preparations used comprise proteins of the retroviral envelope in various forms, for example inactivated viruses, envelope proteins such as the gp 120 and gp 160 proteins of HIV (see in particular GORSE, G. J., Vaccine 10: 383-388, 1992), virus cores with envelope proteins, or envelope proteins associated with various vectors (chimeric viruses, bacteria); see Levy J. A., Trans. Med. Rev. 2: 265-271, 1988 and Microbiol. Rev. 57: 183-289, 1993, in particular page 247.
Other preparations use fragments of the retroviral envelope or immunodominant peptides derived from the envelope glycoproteins, these peptides being presented in various forms (lipopeptides, peptides bound to a supporting protein), so as to make them immunogenic; see in particular Eriksson et al., Vaccine, 11: 859-865 (1993).
The vaccine strategies conventionally described, for example in the field of human, simian or feline AIDS, recommend not modifying the conserved and immunodominant epitopes of the envelope proteins, which may appear to be completely logical. Indeed, on the one hand, these conserved epitopes are common to different viral strains, which is favorable to the production of a vaccine which has to induce an immune response directed against a majority of strains. On the other hand, these immunodominant epitopes are well recognized by the cellular or humoral immune system during the vaccinal and infectious process and, moreover, they frequently represent neutralization sites; see for example HO et al., J. Virol. 61: 2024-2028 (1987); JOHNSON et al., J. Exp. Med. 175: 961-971 (1992); SHAFFERMAN et al., P.N.A.S. U.S.A. 88: 7126-7130 (1991); and HAMMOND et al., J. Immunol. 146: 1470-1477 (1991).
The method of the invention consists, by contrast, in modifying the conserved and immunodominant epitopes of certain proteins of the viral envelope, in order to obtain an effective vaccine. Indeed, the authors of the present invention have discovered that conserved and immunodominant regions of the retroviral envelope may be responsible for harmful autoimmune phenomena. By way of example, in the case of human AIDS, they have observed that certain conserved and immunodominant regions of the HIV envelope exhibit three-dimensional structural analogies and/or cross-reactions with certain regions of at least one protein of the human immune system, such that the administration, as a vaccine, of a viral protein containing said intact regions induces an immune response which is responsible for harmful autoimmune reactions leading to vaccine failure.
At the origin of the present invention, there is, on the one hand, the observation mentioned above that conserved and immunodominant regions of certain retroviruses, usually present in vaccine preparations, are precisely, in a good number of cases, regions which cause harmful autoimmune reactions because they exhibit three-dimensional structural analogies and/or cross-reactions with certain proteins of the host for the virus. At the origin of the present invention, there is also, on the other hand, the observation that said proteins of the host use the same target cell, or the same target cells, as said retroviruses. All these observations carried out by the authors of the invention have led them to think that the retroviral envelope proteins and the host proteins which exhibit three-dimensional structural analogies and/or cross-reactions bind in many cases to the same target cells and possess, on these target cells, common membrane receptors.
It is said that a protein exhibits cross-reactivity with another protein when it is possible to obtain, by in vivo or in vitro immunization with the aid of one of said proteins, an immune response also directed against the other protein, for example when this immunization induces a (so-called B type) humoral response and makes it possible to obtain and to select at least one monoclonal antibody which is capable of recognizing the other protein, or when the same cellular immune response (that is to say of the T type) induced in vitro by one of the proteins recognizes the two proteins, according to the known tests for detecting a T-type immune response, such as for example the tests for cytotoxicity in vitro. It is known that the term xe2x80x9cimmunizationxe2x80x9d denotes the process of induction of an immune response following stimulation, by bringing immunocompetent cells of a host into contact in vivo or in vitro with an antigen, and that one of the aims of the administration of a vaccine agent is precisely to obtain such an immunization.
The subject of the invention is therefore a method of obtaining a vaccine against the pathogenic effects related to the infection of an animal or human host by a retrovirus capable of penetrating into a target cell of said host, said target cell possessing a membrane receptor for a protein of said host, method in which a vaccine agent based on a polypeptide comprising at least part of an envelope protein of a pathogenic strain of said retrovirus is prepared, and in which said polypeptide is prepared in a modified form, it being understood that:
said part of the envelope protein is chosen from those which comprise at least one fragment of an immunodominant region of said envelope protein, said fragment containing at least one amino acid which is a conserved amino acid of said immunodominant region and which is present in said pathogenic strain,
said polypeptide, in the unmodified state, induces an immune response directed both against said immunodominant region and against the protein of the host,
and said modified polypeptide is chosen by [sic] those which induce an immune response directed against said immunodominant region of the envelope protein and not against the protein of the host.
In the definition of the method of the invention which has just been given, the vaccine agent is said to be xe2x80x9cbasedxe2x80x9d on a modified polypeptide. This means that the vaccine agent comprises such a modified polypeptide, but this does not mean that the vaccine agent is necessarily of an exclusively polypeptide nature. In fact, in this vaccine agent, said polypeptide may be optionally bound to (in particular covalently) or associated, in a manner known per se, with any biocompatible molecule which may be chosen, for example from polymers, lipids, peptides (including lipopeptides, glycopeptides, proteins), nucleic acids, oligosaccharides and the like. Said biocompatible molecule may in particular serve as a support for the polypeptide immunogenic agent. It can also serve to modify the conformation of the polypeptide and, in the latter case, said molecule should be considered as a substituent modifying the amino acid residue to which it is attached, said substituent thus modifying, in the final analysis, the antigenicity of the polypeptide of which this amino acid residue is a part.
The method of the invention may comprise, at least in a preliminary research phase, a step consisting in selecting the polypeptides (unmodified) comprising at least part, as defined above, of the viral envelope protein of a pathogenic strain of the retrovirus. This protein part, which comprises at least one immunogenic fragment of an immunodominant region, is such that the polypeptide (unmodified) is capable of inducing an immune response directed both against the viral protein (more precisely against the fragment of the immunodominant region contained in said part) and against the protein of the host, and it is the existence of such an immune response, directed against the viral envelope protein and against the protein of the host, which defines, in the present application, the pathogenic character of a viral strain. It is thus possible to select the polypeptides (unmodified) comprising such a fragment.
A polypeptide fragment is said to be immunogenic if the immunization of a host, in vivo or in vitro, with said fragment, optionally bound to an appropriate support (such as a protein, a lipid or a polypeptide), makes it possible to obtain an immune response (of the B type and/or of the T type, directed against said polypeptide fragment).
In the present application, when reference is made to an immune response, without any other specific information, it is an immune response of a vertebrate, following immunization in vitro or in vivo.
The method of the invention may also comprise at least one step consisting in modifying, in the manner which will be indicated below, a polypeptide thus selected, and in choosing among the polypeptides those modified, at least one modified polypeptide which induces an immune response directed against the viral envelope protein and not against the protein of the host.
Thus, while the prior art taught, as noted above, not to modify the conserved and immunodominant epitopes of the retroviral envelope proteins, the aim of the method of the invention is, by contrast, to modify the antigenicity of such epitopes so as to obtain a differential immune response with respect to the viral envelope protein and to a protein of the host.
It is known that in order to modify the antigenicity of an immunogenic fragment of a polypeptide, it is possible to modify said polypeptide with the aid of a mutation affecting at least one amino acid. A definition will be given later of what xe2x80x9cmutationxe2x80x9d should be understood to mean here. The mutated amino acid may be present in the immunogenic fragment, or even in a region of the polypeptide outside said fragment. It is in fact known that the modification of an amino acid situated outside a fragment can affect the spatial structure of said fragment and therefore its antigenicity; in particular, it has been shown that the conformation of an amino acid residue, in a peptide, can be influenced by the nature of the amino acid residues at positions going from +8 to xe2x88x928 relative to this amino acid residue; see for example GARNIER et al., J. Mol. Biol. 120: 97-120 (1978). Beyond this, the nature of the amino acid residues still has an influence, but this influence is neither symmetrical nor quantifiable from the sole knowledge of the peptide sequence considered.
A mutated amino acid can therefore be situated in the modified polypeptide, inside or outside the immunogenic fragment. When it is outside the immunogenic fragment, it is generally not separated from the nearest end of said immunogenic fragment, in the polypeptide chain, by more than eight (and in particular by more than seven) amino acid residues. In particular, an amino acid, mutated in accordance with the present invention, and situated outside the immunogenic fragment, is generally not separated by more than eight amino acid residues, and in particular by more than seven amino acid residues, from the nearest conserved amino acid belonging to the immunodominant region of which at least one fragment is contained in the unmodified polypeptide.
The modified polypeptide in accordance with the present invention may be, for example, the whole envelope protein of a pathogenic viral strain, modified by at least one mutation as indicated above. The modified polypeptide may also be part of the envelope protein of a pathogenic viral strain, modified by at least one mutation as indicated above, said part comprising at least one immunogenic fragment as defined above. The modified polypeptide may also be a chimeric protein comprising at least part of the envelope protein, said part of the envelope protein being as defined above and comprising at least one mutation.
The modified polypeptide used according to the invention may be, for example, a transmembrane glycoprotein of a retrovirus or a fragment of a transmembrane glycoprotein, in particular a fragment comprising an outer region of said transmembrane glycoprotein (modified), that is to say a region which is present on the outer surface of the viral membrane. Of course, such a protein fragment comprises at least part of an immunodominant region, as indicated above. When reference is made to an xe2x80x9couterxe2x80x9d region of a protein, it is more precisely its surface which is accessible to the solvent, which may be defined in particular with the aid of software such as X-plor (see below) using the algorithm described by Lee and Richards, J. Mol. Biol. 55: 379-400, 1971. Said polypeptide may also be in the form of an oligomer of at least part of said transmembrane glycoprotein, in the modified state.
The definition given above of the method of the invention implies that the polypeptide used comprises at least part of an immunodominant and conserved region of a viral envelope protein. In the description of the present application, xe2x80x9cconserved regionxe2x80x9d describes a region, optionally reduced to a single amino acid residue, of the viral protein, where, for a majority of strains of a given virus (for example in at least 6 strains out of 10 approximately), there are one or more identical or functionally analogous amino acids situated at the same position in peptide sequence alignments of said protein of the various strains. Such an identical or functionally analogous amino acid is called conserved amino acid. The notion of conservation of functionally analogous amino acids is known, and there are numerous substitution matrices which make it possible to quantify this notion (Dayhoff, M. O. et al., in Atlas of Protein Sequence and Structure, 1978, vol. 5, Suppl. 3, Chapters 22 and 23).
The conserved regions can be easily determined, after sequencing of proteins of various strains of the viruses studied, by methods of multiple alignments of the sequences obtained. For that, it is possible to use, for example, the Clustal-w program (Thompson, J. D. et al., Nucleic Acids Research 22: 4673-4680, 1994). Moreover, the protein sequences of various viral strains are often accessible on data banks. For example, the Web server of the Los Alamos HIV data base has the HIV 1, HIV2, SIV and FIV sequences which are regularly updated.
The appended Tables 2a, 2b, 2c and 2d are examples of sequence alignment of the regions belonging to the homologous envelope glycoproteins of region 545-682 of the HIV transmembrane glycoprotein (entry SWISSPROT ENV_HV1 BR), respectively for HIV1, HIV2, FIV and SIV.
Table 2a shows the sequences represented by SEQ ID NOs: 1-29 in number order from top to bottom. Table 2b shows the sequences represented by SEQ ID NOs: 30-39 in number order from top to bottom. Table 2c shows the sequences represented by SEQ ID NOs: 40-46 in number order from top to bottom. Table 2d shows the sequences represented by SEQ ID NOs: 47-57 in number order from top to bottom. The last line of the tables summarizes, with the aid of symbols, the degree of homology and therefore the degree of conservation observed. The symbol xe2x80x9c*xe2x80x9d indicates a position of the alignment where the same residue is present in all the sequences, the symbol xe2x80x9c:xe2x80x9d indicates a position in the alignment where the amino acids present in the various sequences are very similar, the symbol xe2x80x9c.xe2x80x9d indicates a position in the alignment where the amino acids present in the various sequences are similar, and the absence of a symbol indicates a position in the alignment where the amino acids present in the various sequences are not very similar. This symbolic system is used by the Clustal W alignment program (version 1.7).
In the description of the present application, immunodominant region of a protein refers to a peptide sequence which induces, in a great majority of cases (for example in at least 7 cases out of 10 approximately), a humoral and/or cellular response of the immune system directed against said region after immunization with a protein containing said sequence or with a peptide essentially consisting of said sequence.
The definition of the method of the invention makes reference to the target cells of a virus which are the cells into which the virus is capable of penetrating. The target cells of retroviruses are generally known. Viruses have the property of binding to the cells which they are capable of infecting. It is therefore optionally possible to test for, using routine experiments in vitro, the target cells of a virus studied.
The definition of the method of the invention also makes reference to the cells of the host having a membrane receptor for a protein of the host. The cells of the host which have a receptor for a protein of said host are often known and, in the opposite case, it is possible, using routine experiments, to determine if a given protein binds to a certain type of cell. It is possible, for example, to use a radiolabelled protein and to determine if it binds to said cell type. It is also possible to test if the protein binds to a given membrane receptor using a cell line transfected with a gene expressing said membrane receptor.
The proteins of the host for which certain cells of the host possess a membrane receptor are mainly proteins belonging to the range of soluble protein mediators. This range includes proteins called, depending on the cases, hormones, growth factors or cytokines, although there is no distinct boundary between these various categories of mediators; see for example CAVAILLON J. M., Les Cytokines (Masson, Paris, 1996) Chapter 1, pages 1-3 and preface.
In the present application, it is considered that an immune response, for example an antibody response, obtained by immunization with the aid of the modified polypeptide prepared in accordance with the method of the invention, is directed against the viral envelope protein and not against the protein of the host, when the antibodies obtained have affinities for the protein of the host and for the envelope protein of the retrovirus which exhibit a substantial difference, resulting in particular in differences in reactivity which are considered to be very significant in ELISA tests, such as for example optical densities in a ratio of about 4 (or more), which means that the optical density observed after attachment of said antibodies to the viral protein is at least four times higher than that observed for the attachment of said antibodies to the protein of the host. Similarly, a cellular type immune response is considered to be directed against the envelope protein but not against the protein of the host when the immunization in vitro of immunocompetent cells of the host with the candidate vaccine induces the formation of activated cells whose reaction toward cells (including transfected cell lines) expressing the retroviral envelope protein is significantly higher than the reaction toward cells expressing the protein of the host, for example when, in the final optical measurement, or in the final radioactivity counting (in particular 51Cr radioactivity released by target cells) of the test used, or alternatively in the assessment by any known means of a cell lysis caused by induced cytotoxic cells, the scales of response are in a ratio of about 4 (or more). The criteria which have just been indicated make it possible at least to make a first choice among the modified peptides studied, but in the final analysis, it is the absence or the decrease in the pathogenic effect due to the suppression or the weakening (demonstrated by any appropriate means) of the immune response toward the protein of the host which will constitute the criterion for selection of the modified peptides capable of constituting satisfactory vaccine agents.
The immunodominant and conserved regions of which it is desired to modify the antigenicity, in accordance with the invention, may be chosen from those which give in vitro a cross-reaction, of the B type and/or of the T type, with the host protein defined above.
It is also possible to choose such an immunodominant and conserved region from those for which a three-dimensional structural analogy with a region of said protein of the host has been determined beforehand, said structural analogy being capable of being associated with a cross-reaction in vitro and/or in vivo. The three-dimensional structural analogy between certain regions of two proteins refers to equivalent arrangements, in space, of amino acid residues which are similar because, in particular, of their side chain and/or of their analogous functional chemical groups. The three-dimensional structures of the proteins can be obtained with the aid of nuclear magnetic resonance (NMR) spectra and/or of X-ray diffraction spectra. For example, the structure of the SIV gp41 protein was obtained with the aid of the NMR spectrum (Caffrey M. et al., J. Mol. Biol. 271, 819-826, 1997). In addition, it is possible, in some cases, to obtain a good model with the aid of molecular modeling techniques, from the atomic coordinates of a protein of known structure. It is possible to use for that, in particular, the molecular modeling software X-plor (reference: xe2x80x9cA system for X-ray crystallography and NMR, Version 3.1xe2x80x9d, Axel T. Brunger, Yale University Press, 1992).
To search for a three-dimensional structural analogy, it is possible to use, for example, the known methods of visualization and superposition on a graphic screen of the three-dimensional structure of biological molecules. Software exists which allows the visualization of the three-dimensional structures of the molecules with different modes of representation, the calculation of the geometric parameters (such as distances, angles and the like) and the objective and quantitative superposition of several molecular structures (in particular RASMOL software: Sayle, R. A. and Milner-White E. J., J. Mol. Biol., 247, 536-540, 1995 and ANTHEPROT software: Geourjon C. and Delxc3xa9age G., J. Mol. Graph. 13, 209-212, 1995) as well as the estimation of the accessibility to solvents (X-plor software, already mentioned, and CCP4 software: Collaborative Computational Project Number 4, Acta Cryst., D50, 760-763, 1994.
However, in order to have a finer estimation of these structural analogies, it is useful to consider, at the level of each amino acid, the functional groups positioned in a similar manner in both proteins which are compared. For that, the co-inventors of the present invention use methods which make it possible to calculate molecular surface areas with the aim of comparing functional properties between two three-dimensional structures, in order to take into account, not amino acids in their entirety, but also, more particularly, functional chemical groups of each of them (for example: amide, carboxyl, hydroxyl, sulfhydryl and amine functions and the like). It is thus possible to take into consideration, in the structures compared, functionally analogous amino acids, and not only identical amino acids.
It is therefore considered that a region of a retroviral protein exhibits a three-dimensional structural analogy with a given region of a protein of the host when the techniques which have just been mentioned make it possible to demonstrate, in the two regions compared, a similar spatial organization of certain identical or functionally analogous amino acids.
It should be noted that amino acids which are functionally analogous and grouped together in a similar manner in space can be relatively distant from each other in the same peptide chain. However, the three-dimensional structural analogy between two proteins which are being compared can also relate to the spatial arrangement, in a similar manner, of identical or functionally analogous amino acids in the case where, one of the proteins being oligomerized, the amino acid residues involved are situated on different chains of the oligomer, whereas the amino acid residues of the other protein which are involved in this analogy can be situated on the same peptide chain of this other protein.
It is particularly advisable to search for three-dimensional structural analogies and/or cross-reactions with regions of the protein of the host which are involved in the attachment of said protein to its receptor.
Among the proteins of the host which are mentioned in the definition of the method of the invention, there may be mentioned in particular the soluble mediators as defined above. Taking into account the remark made above that immunosuppressive effects are generally associated with retroviral infections, it is particularly important to search for structural analogies and/or for cross-reactions between an outer protein of a retrovirus and soluble protein mediators of the immune system. Among these immune system mediators, there may be mentioned cytokines, and in particular interleukin-2, interleukin-10, interleukin-15 as well as interleukin-8 and chemokines.
To prepare the modified polypeptide which constitutes the vaccine agent obtained according to the invention, it is possible to use known methods of peptide synthesis or genetic engineering techniques. It is possible to isolate or to prepare a polynucleotide sequence encoding at least part of the envelope protein of the virus and, if desired, it is possible to introduce at this stage, into the nucleotide sequence, mutations which make it possible to obtain a mutated product of translation which constitutes the modified polypeptide. It is also possible to directly synthesize a modified polynucleotide sequence comprising one or more mutations and encoding the modified polypeptide. The mutated polynucleotide sequences thus obtained are introduced in a known manner into an appropriate vector which makes it possible to express said polypeptide, optionally in modified form. Such a vector is for example E. coli, a baculovirus or a mammalian cell. It is also possible to carry out the mutation on an unmodified polypeptide obtained according to one of the preceding methods.
In the present application, xe2x80x9cmutationxe2x80x9d refers to any modification of a region (optionally reduced to a single amino acid residue) of a polypeptide, by physical means, chemical means (covalent or noncovalent modification) and/or biological means (mutations by substitution, deletion and/or insertion of one or more amino acids), leading to the modification of the functional potentials of the constituent amino acid(s) of said region, termed xe2x80x9cmutated regionxe2x80x9d. By way of example, it is possible to carry out mutations leading to the abolition, acquisition and/or modulation of the properties of disulfide bridges, hydrogen bonds, electrostatic interactions and/or hydrophobic interactions, the modification of the capacity of a protein to form a heterocomplex, or alternatively, in the case of an oligomeric protein, the modification of the state of oligomerization or of the stability of the oligomer.
The modification of an amino acid a of a polypeptide chain (including the modification of a terminal amino acid of the polypeptide considered) can influence the conformation of the neighboring amino acids in the chain, including, as was recalled above, the conformation of an amino acid b separated from a by a number of amino acid residues which may be as high as seven or eight, and when the amino acid b is part of an epitope, any modification of the amino acid a (in particular any addition of a substituent or any modification of a substituent) is capable of modifying the antigenicity of the epitope considered.
In the phase for searching for modified polypeptides in accordance with the invention, the choice of the amino acids to be mutated and/or the choice of the mutation methods can be made in an arbitrary manner or in a reasoned manner. It is possible to use in particular at least one of the following methods of modification:
1) the replacement of one or more amino acids having a hydrophobic side chain (examples: Ala, Leu, Val, Ile, Phe, Trp, Met, Tyr, Cys) by one or more amino acids having a hydrophilic side chain (examples: Arg, Asp, Asn, Glu, Gln, Ser, Thr, His, Lys) or an indifferent chain (examples: Gly, Pro) and vice versa;
2) the replacement of one or more amino acids having a positively charged side chain (examples: Arg, Lys, His) by one or more amino acids having a negatively charged side chain (examples: Asp, Glu) or a neutral chain and vice versa;
3) the acquisition, suppression and/or modification of one or more disulfide bridges;
4) the production of mimotopes, in particular which are obtained by retro-inversion;
5) the substitutions, suppressions, additions and/or other modifications of at least one amino acid which is potentially a donor or acceptor of hydrogen bonds;
6) the substitutions, suppressions, additions and/or other modifications of at least one amino acid which is potentially a donor or acceptor of ionic bonds;
7) the change in steric hindrance by substitutions, suppressions, additions and/or other modifications of one or more amino acids;
8) the use of amino acids which are not naturally present in proteins;
9) the modification of glycosylation (creation, suppression or modification of glycosylation sites or of their associated sugars).
Of course, the modified polypeptides thus obtained are tested, as indicated above, in order to select the modified polypeptides which induce an immune response directed against the envelope protein and not against the protein of the host.
The method of the invention may be applied to the obtaining of vaccines, in particular against the following viruses: HIV, FIV, SIV, leukomogenic oncoviruses (avian, murine, feline, human, simian and bovine leukemia viruses, that is to say, respectively, ALV, MULV, FELV, HTLV, STLV, BLV), primate type D retroviruses, mammary tumor-inducing type B retroviruses, Rous sarcoma virus, maedi-visna virus (infecting sheep), feline sarcoma virus, avian myelocytomatosis virus and avian myeloblastosis virus.
The subject of the invention is also the use of a modified polypeptide, as defined above, in the preparation of a vaccine composition for preventing the pathogenic effects related to the infection of a host by a retrovirus.
The invention also relates to a vaccine composition which can be obtained by the method of the invention, and containing as active ingredient a modified polypeptide as described above. Such a composition may be used in a method of vaccination for preventing the pathogenic effects of retrovirus infections, this method essentially consisting in administering to a vertebrate animal, including a human, a modified polypeptide as defined above in a sufficient quantity to obtain a vaccination effect. The formulation of the vaccine compositions, and their method of administration, are known per se and will not be further described here.
The subject of the invention is also a modified retroviral polynucleotide encoding a modified polypeptide as defined above. The modified polynucleotide may be obtained as indicated above. The invention extends to an expression vector into which said modified polynucleotide has been inserted, said expression vector being thus capable of expressing the modified polypeptide.
The modified polypeptide obtained according to the invention can also serve as immunogenic agent in order to induce, by immunization, the formation of antibodies which can be used in particular in the treatment of retroviral infections, and the invention therefore also relates to the antibodies obtained in response to the immunization of animals (including humans), in vivo or in vitro, with the aid of the vaccine agent containing a modified polypeptide described above. The antibodies of the invention are in particular purified polyclonal antibodies or monoclonal antibodies exhibiting the characteristic of recognizing the retroviral envelope protein without recognizing the protein of the host. The purification of the polyclonal antibodies, and the selection of the monoclonal antibodies, can be carried out with the aid of the viral protein and of the protein of the host, so as to select only the antibodies which recognize the viral protein and not the protein of the host. The antibodies of the invention can be used in particular in the early treatment of infections caused in said host by the retrovirus against which they are directed. The dosage is the usual dosage for antibodies. The pharmaceutical compositions containing such antibodies also constitute one of the subjects of the invention.

Method for obtaining vaccines for preventing the pathogenic effects related to a retroviral infection


The animal stun gun of the invention is a non-lethal alternative to subduing a dog or other animal in an imminent attack situation where, previously, the use of firearms was the only alternative to stop its advance. The stun gun is an electrical self-defense device that uses high voltage to stop the attacking dog or animal by touching it with the prongs of its staff in turning on an electrical pulse generator to immobilize the dog or animal with a very low amperage, high voltage pulse of high frequency to make its muscles work rapidly, although not efficiently.

This invention relates to stun guns for animals, in general, and to such stun guns for use against vicious and wild dogs, in particular.
Stun guns that temporarily immobilize humans are in lawful use by law enforcement agencies in many parts of the United States. Recently, it has become apparent that a similar nonlethal alternative would be beneficial to subdue a vicious or wild dog, in an imminent attack situation. This has become all the more so with the increasing use by drug dealers of pit bull terriers, rottweilers and german shepherds as protectors and bodyguards in their criminal operations. As will be understood by those involved in police law administration, such dogs continue to attack, even after being initially, although not mortally, shot. Since many of the drug enterprises raided are carried on in an apartment setting, shooting in close quarters is preferably to be avoided. The understanding thus becomes clear that a non-lethal alternative is desirablexe2x80x94however, one which continues to be fully protective of the law enforcement personnel carrying out their sworn duties.
It is an object of the present invention, therefore, to provide an animal stun gun as this non-lethal alternative.
It is an object of the invention, also, to provide such an animal stun gun which can continue to be used effectively, even in the event that its operative nature should somehow fail.
It is another object of the present invention to provide an animal stun gun of this type which only temporarily immobilizes the animal, so as not to cause it permanent injury.
As will become clear from the following description, the animal stun gun of the invention is essentially an electrical self-defense device that employs high voltages to stop an attacking dog, although it could be used for other animals, as well. As will be seen, touching the dog with the prongs on the staff employed quickly immobilizes it, to prevent or impede any ensuing attack. However, because the amperage involved is selected quite low, serious or permanent injury is avoided. In particular, the stun gun of the invention is designed to key into the nervous systemxe2x80x94depositing its energy into the muscles of the dog at a high pulse frequency which causes them to work very rapidly, although not efficiently. Such rapid work cycle will be understood to deplete blood sugar by converting it into lactic acid in only a matter of seconds. The energy loss which results will be recognized as making it difficult, for the dog to move and function. At the same time, the tiny neurological impulses which otherwise travel throughout the body in directing muscle movement are interruptedxe2x80x94thereby causing a deterioration and loss of balance which puts the dog in a passive and confused condition for several minutes. Still, on the other hand, there will be seen to be no significant effect on the dog""s heart or other organs.
Specifically, and in accordance with the present invention, a preferred embodiment includes a staff of a length at least 3 feet to keep the dog away from the holder of the stun gun, along with a power source housed with the staff capable of providing pulses of electrical energy of at least one hundred thousand volts at a current of at least 2 milliamperes. This stun gun embodiment also will be seen to employ a pair of metal prongs at a first end of the staff to receive these pulses of electrical energy, with a sleeve coupled with the staff to quiescently overlay and shield the metal prongs, yet slidable rearwardly to expose them. In accordance with the invention, a switch operates with the sleeve to turn on the power source for delivering the pulses of electrical energy to the prongs, with the switch being arranged to automatically turn on the power source once the overlying sleeve is slid rearwardly a predetermined amount.
In the preferred embodiment of the invention to be described, the sleeve is moved rearwardly of the metal prongs by the force exerted by the skin of the dog against which the front end of the staff is pressed. Initially overlying the prongs by some 1 inch or so, the sleeve is spring-biased to turn off the power source and to shield the metal prongs once again upon the release of the pressure of the dog""s skin. In the embodiment to be described, the power source employed is capable of providing these pulses of electrical energy at a current of substantially 3 milliamperes, within a range of three hundred-four hundred thousand volts, for a staff substantially 5 feet in length.
While any appropriate electrical power pulsing source could be employed, in accordance with the invention, such source could be included within the staff, itselfxe2x80x94as when the staff is formed as a tubular polexe2x80x94or secured to it if not so hollowed out. In further accordance with the invention, the stun gun could also employ a tightenable noose large enough to loop about the neck of the immobilized dog, for securing it to drag it to a safe location.
In the specific embodiment described below, the metal prongs employed at the forward end of the staff are selected of a xe2x80x9cbarbedxe2x80x9d configuration, to guide and push the dog away, in the direction in which it is to be pointed until the electrical pulses take effect. At the same time, such xe2x80x9cbarbsxe2x80x9d effectively serve as a xe2x80x9cstopxe2x80x9d to limit the extent of penetration of the prongs into the skin, thereby affording a degree of protection to the dog itself. The prongs, in this embodiment, are constructed of spring-steel (for example), and the switch which automatically turns on the power source once the sleeve is slid rearwardly the required amount could be fabricated as a magnetically controlled reed switch.
With the invention as thus set out, as a general rule, only a one-half second contact would be required to repel and startle the dog, producing some degree of pain and muscle contraction. Holding the prongs against the dog for a one-to-two second interval causes a degree of muscle spasm and a dazed mental state. Holding the prongs while pulse energizing over three seconds causes a loss of balance and muscle control, along with a significant amount of mental confusion and disorientation. In such circumstances, the noose could then be looped over the dog""s neck to gain further control of itxe2x80x94all in an environment which militates against the need for having to shoot the dog until it stopped coming.
As will be appreciated by those involved in the field of police law enforcement, the bringing of a stun gun of this type to the scene generally follows from intelligence gathered from undercover personnel or from a confidential informant that at the xe2x80x9cdrugxe2x80x9d scene, for example, such a violent protector dog is maintained.

Pulse-Wave Protective Devices International, Inc.


Residuelessly redetachable contact-adhesive sheet-like structures wherein are provided approximately calotte-shaped bonding sites which are up to about 600 xcexcm in diameter at the base where the calottes stick to the sheet-like structure and which are produced by screen or gravure printing using high-solids dispersion of adhesive substance.

The present invention relates to contact-adhesive sheetlike structures which, after attachment, can be residuelessly redetached and, if desired, attached again. Sheet-like structures of this type have long been used in practice, examples thereof being residuelessly detachable reversibly adherent contact-adhesive labels, protective films, masking papers, note sheets and advertising materials and residuelessly detachable and reattachable cohesive plasters and dressings.
A number of different contact-adhesive materials and manufacturing methods have already been described for these purposes.
German Offenlegungsschrift No. 2,407,494 describes a weakly contact-adhesive emulsion polymer which is prepared by copolymerizing 2-ethylhexyl acrylate, methyl methacrylate, (meth)acrylic acid and 1,4-butanediol diacrylate in the presence of plasticizers such as paraffin oil, squalene or low molecular weight polyisobutylene and, owing to its relatively low adhesivity, permits redetachability of the contact-adhesive articles equipped therewith.
Japanese Pat. No. 82/87,481 describes an aqueous contact-adhesive dispersion which is prepared by copolymerizing esters of acrylic acid with alkyl radicals of 4 or more carbon atoms, methacrylic acid and N-methylolacrylamide in the presence of special plasticizers. After addition of thickening agents these dispersions are suitable for equipping reversibly adherent paper labels. Japanese Pat. No. 82/70,162 describes a similar process.
Japanese Pat. No. 82/31,972 describes an emulsion copolymer which is prepared from 2-ethylhexyl acrylate, methyl methacrylate, acrylic acid, itaconic acid and ethylene dimethacrylate and, after buffering with sodium acetate and thickening with hydroxyethyl cellulose, is suitable as a contact adhesive for redetachable contact adhesive articles.
Japanese Pat. No. 82/42,778 describes a weakly contact-adhesive bead copolymer which is prepared in aqueous dispersion from (meth)acrylic esters in the presence of a dispersant. Said water-soluble dispersant consists of a copolymer which is obtained by free-radical copolymerization from a low proportion of butyl acrylate and a high proportion of acrylic acid.
U.S. Pat. No. 3,691,140 and German Offenlegungsschrift No. 2,417,312 describe the production of redetachable, repeatedly usable contact-adhesive articles whose contact-adhesive layer consists of contact-adhesive microspheres. Said microspheres are synthesized in aqueous dispersion in the presence of anionic emulsifiers by copolymerizing alkyl acrylates, special water-soluble ionic monomers and maleic anhydride. To improve the anchoring, the microspheres are affixed to the carrier materials by means of special anchoring layers.
In U.S. Pat. No. 2,510,120, the redetachability of contact-adhesive sheetlike structures is improved by coating part of the surface, for example in the form of strips or other two-dimensional patterns.
German Offenlegungsschrift No. 2,535,897 describes a part-surface application of contact-adhesive compositions to label paper by using an engraved roll to coat only the label area in full before the punching out.
Contact-adhesive articles which are equipped with suitable contact-adhesive compositions by the cited methods have not only advantageous properties but also serious shortcomings.
1. Plasticizers, which are frequently contained in the contact-adhesive compositions described, for example paraffin oils, tend to bleed out of the composition and can stain the carrier materials, for example paper. This also affects the adhesive properties. Finally, on prolonged attachment they can also penetrate into the attached substrates.
2. Thickeners and emulsifiers settle out on the contact-adhesive particles as the emulsion dries and forms a film; they reduce the adhesivity and promote, in particular, the absorption of water by the contact-adhesive films in dependence upon the ambient humidity and the moisture content of the attached substrates. The plasticizing action of even small amounts of water affects the adhesion properties in an uncontrollable manner; this impairs the usability of contact-adhesive articles of this type.
3. The adhesivity of a whole-surface coat of contact adhesive increases with the duration of attachment, since, to form a bond, films of contact adhesive must possess some flow, albeit limited, because without that property the substrate on which attachment is to take place cannot be wetted. In the case of short-lived attachment, only the protuberances of the substrate surface are wetted, and the strength of attachment is correspondingly low. As the duration of attachment increases, the contact adhesive flows also into the indentations of the substrate surface, which is normally not microscopically smooth. As a result, the force required to separate the attached surfaces rises to two to three times the starting values. As a consequence, the carrier material or the contact adhesive film can split (cohesive failure) on breaking the attachment. The substrate is soiled, and the contact-adhesive article is unsuitable for reuse. In some instances, moreover, the surfaces of less strong substrates, for example papers, can be damaged. As contact-adhesive articles are used on substrates of all kinds of roughness, strength and chemical composition, a satisfactory match between adequate initial adhesivity and reversibly rebreakable end adhesivity is possible for whole-surface smooth coatings only in specific cases.
4. Part-surface coatings of contact adhesive which are applied to the carrier materials in the form of strips or other patterns can be advantageous, in particular for carriers having lower tear strength, such as, for example, paper. Owing to the smaller area of attachment, the tensile forces exerted on the carrier material in the course of separating the attached surfaces are kept in check, and splitting of the carrier material is avoided. However, in the areas of whole-surface attachment the force of adhesion to the substrate increases with time by the mechanism described in the preceding section, so that on suitably long attachment there is a danger that the contact-adhesive film will split on separation of the attached surfaces, in particular when the attached surfaces are separated in the direction of the strips.
5. Coatings of contact-adhesive microspheres of a suitable size (50-150 xcexcm) show the time-dependent increase in the strength of attachment to a small extent. The elastomeric recovery forces of the spheres flattened off in the course of the pressure-sensitive attachment to the substrate tend toward the re-formation of the originally curved surface and thereby counteract any stronger flow of the contact adhesive into the microinch indentations of the substrate. The force employed in separating the attached surfaces acts concentratedly on the relatively small, circular, discrete attached flattened parts of the spheres and is distributed from there into the much bigger sphere volumes. As a result, the desired adhesive failure is brought about preferentially, and the undesirable cohesive failure is suppressed. Besides these advantages, this process also has some shortcomings and/or limitations. Anchoring the microspheres to the carrier requires a binder layer in which the spheres are partly submerged and which, owing to the larger wetting area, effects anchoring to the carrier surface. However, this principle requires a flat surface of limited absorbency; in order to prevent the binder from penetrating into the substrate and thus becoming ineffective in the sense described above. The choice of the carriers which are coatable by this principle is limited as a result and/or necessitates an additional, smoothing coat, for example for paper.
Moreover, application of the microspheres to the carrier material must be done in such a way as to produce a monoparticulate coating. In areas where microspheres are positioned on top of one another, reliable anchorage is no longer assured, since the anchoring mechanism described above then becomes ineffective. This condition is difficult to meet with a statistically clearly measurable size distribution of the microspheres. The statistical size distribution of the contact-adhesive microspheres, which matches that of a Gaussian distribution function, also has the disadvantage that the larger microspheres which protrude farther above the carrier surface are particularly highly stressed in pressure-sensitive adhesion, while the smaller ones are under little stress, if any. Furthermore, the recovery forces of the larger spheres have the effect that the bonds with the smaller spheres which are formed under the contact pressure break with time and cause a reduction in the strength of the bond.
It is the object of the present invention as far as possible to avoid the disadvantages described.
This object is achieved, surprisingly, with contact-adhesive calottes of defined, uniform size which are applied to suitable carrier materials in predetermined spacing and which are found to have excellent adhesive properties for manufacturing reversibly adherent, residuelessly detachable sheetlike structures. By means of their flat bottom surfaces, contact-adhesive calottes give more reliable anchorage to the carrier materials than, for example, corresponding microspheres do. Moreover, this generally requires no additional anchoring layers. The spherical surfaces of the calottes face the direction of the substrate to which they are to be attached and offer the contact-adhesion advantages which curved, contact-adhesive surfaces have in respect of reversible, residuelessly breakable attachment and which have already been described in section 5.
The invention accordingly provides residuelessly redetachable contact-adhesive sheetlike structures wherein as provided approximately calotte-shaped bonding sites which are up to about 600 xcexcm in diameter at the base where the calottes stick to the sheetlike structure and which are produced by screen or gravure printing using high-solids dispersions of adhesive substances.
These reversibly adherent contact adhesive articles are favorably distinguished from those of the state of the art and, in addition to having convincing advantages, have the state of the art disadvantages to a much smaller degree.
The desired calotte shape of the bonding sites need not be fully realized in practice. For the purposes of the invention it is thus possible to use bonding sites having a non-circular or not perfectly circular base, since, owing to the pronounced structural viscosity and thixotropy, the bonding site assumes an approximate calotte shape. The adhesive composition used, accordingly, preferably has on application a pronounced structural viscosity and thixotropy, sufficiently high for forming bonding sites of approximate calotte shape.
Further preferred features of the adhesive composition are good contact-adhesivity, good film-forming properties and adequate elastomeric properties.
Suitable contact adhesives are in particular those based on (meth)acrylic acid esters with alkyl radicals of 4 to 12 carbon atoms.
However, small amounts of (meth)acrylic acid esters with alkyl radicals of 1 to 3 carbon atoms or of 13 to 18 carbon atoms can also be present. Small amounts (about 0-12%) of (meth)acrylic acid and/or other copolymerizable acids, such as maleic acid, fumaric acid or itaconic acid, can be present as copolymerized units. To increase the cohesion and improve the stability of the dispersion, it is also possible to use acrylonitrile or acrylamide and added crosslinking agents, for example N-methylolacrylamide or glycidyl methacrylate in conjunction with hydroxyl-carrying (meth)acrylic acid esters or polyfunctional acrylates, for example butanediol bisacrylate. Finally, some of the (meth)acrylic acid ester can be replaced by copolymerizable compounds such as vinyl acetate or vinyl propionate.
Contact-adhesive calottes anchored at their base to carrying materials are best produced by means of industrial printing methods of the screen-printing or gravure-printing type which are known per se, using, as the contact adhesives, dispersions of high solids content, preferably concentrated thixotropic aqueous contact-adhesive dispersions. Aqueous dispersions are preferred, but it is also possible to use dispersions of the organosol type, i.e. dispersions based on a high-boiling organic non-solvent, or even of the plastisol type, such as pastelike products comprising plasticizer plus plastics material. Aqueous dispersions preferably have a solids content of at least 45% by weight, especially of about 55-65% by weight.
Rotary screen printing basically comprises using a rotating seamless drum-shaped perforated round screen. In the inner shell, a mechanically or magnetically supported round or four-edged doctor squeezes the contact-adhesive dispersion fed into the drum onto the carrier web through the perforations of the screen wall. The carrier web is guided along the outside shell of the screen drum by means of a back-pressure roller at a speed which corresponds to the circumferential speed of the rotating screen drum.
The coating is subsequently dried in a hot-air duct or by infrared, i.e. high-frequency radiation.
The calotte shape is formed in this process by the following mechanism: the pressure from the doctor blade feeds the contact-adhesive dispersion through the perforations of the screen onto the carrier material. This carrier material is first of all surface-wetted by the dispersion in line with the hole geometry. The size of the calotte base thus formed is predetermined by the diameter of the hole in the screen. The bore is lifted away from the carrier as a function of the transport speed of the carrier web, i.e. as a function of the speed of rotation of the screen drum. Due to the internal cohesiveness of the contact-adhesive dispersion, the supply of contact-adhesive dispersion stored in the bore is drawn down by the base which is already adhering to the carrier, that is to say pushed by the doctor pressure onto the web. At the end of this transport of material, the more or less highly curved surface of the calotte forms above the predetermined base area, depending on the structural viscosity, thixotropy and flow of the dispersion. The ratio of height to base of the calotte, in line with this mechanism, substantially depends on the ratio of hole diameter to wall thickness of the screen drum and the physical properties (flow behavior, surface tension, angle of contact of the carrier material) of the dispersion. On drying, the height of the calotte decreases in line with the loss of water. The size and shape of the base which becomes anchored to the carrier material in the course of drying changes very little, if at all.
The calotte formation mechanism described requires carrier materials which are absorbent or at least wettable by the adhesive compositions. Non-wettable carrier surfaces should be treated by additional measures, such as, for example, electric corona discharge or coating with wettability-conferring substances.
The size and shape of the calottes can be definitively predetermined with the printing method described. Differences in size which are relevant to application and which, as described, can impair the quality of contact adhesive articles do not arise if this coating method is carried out correctly. The base diameter of the calottes can be chosen within the range from about 30 xcexcm to about 600 xcexcm) and the height of the calottes correspondingly from about 30 xcexcm to about 600 xcexcm. Base diameters of 80-500 xcexcm are preferred, smaller diameters being suitable for smooth carrier materials and larger diameters, with correspondingly greater calotte height, for rougher or highly porous carrier materials.
The positioning of the calottes on the carrier is definitively predetermined by the geometry of the application system which is variable within wide limits, for example screen geometry or gravure geometry. The adjustable parameters refered to allow the desired adhesion properties of the coating to be adjusted very accurately, for matching to the various carrier materials and end uses.
The sheetlike structures according to the invention featuring mechanically applied contact-adhesive hemispheres, possess inter alia the advantages of the known microspheres without having their disadvantages:
It is found that mechanically applied calottes have a significantly narrower size distribution than chemically produced bead polymers. Whereas polymerization reactions produce molecular sizes or bead polymer sizes which follow a Gaussian distribution curve, tools for the mechanical application of calottes can be manufactured within tolerances tending toward zero.
It is also found that all applied calottes are involved in the bonding, since, owing to the manner of application, they all have the same dimensions, so that only the actually necessary amount of adhesive need be made available. This has the additional benefit of an appreciable saving of adhesive. Moreover, the adhesive bond is improved compared with state of the art reversible adhesive bonds.
It is also found that every carrier can be coated, irrespective of its surface roughness and absorbency, extremely economically. In particular, there is no need to smooth the carrier surface by mechanical action, such as, for example, glazing, or to apply preliminary coatings to fill out any unevenness. The disappearance of adhesive dispersions into porous carrier material can be avoided by using particularly coarsely divided dispersions. However, if the carrier material has poor adhesion properties, such as, for example, polyethylene, it is possible to use a primer which then has to act as a tackifier.
It is also found that reliable adhesion requires no binder, since the anchoring is ensured by the base area which is adapted to the surface structure of the carrier and which at any rate is larger than the area facing the substrate and involved in the bonding there.
It is also found that the size of the calottes is variable within very wide limits without the effect of the reversible attachment option being lost.
It is also found that the properties of the substrates contact-adhesively coated in this way can be adjusted for almost any purpose in which reversibility is required through the freely selectable parameters of the diameter of the calottes at the base, height of the calottes, geometric distribution of the calottes, and frequency (number per unit area).
It is also found that the properties of the adhesive material can be modified within a very wide range: from very soft, highly tacky and of low shearing strength to hard, slightly tacky and of high shearing strength.
It is also found that the three-dimensional shape of the calottes or approximate calottes, with its advantageous properties, is stable in reversible attachment due to its elastomeric properties. If only for that reason, this shape is constantly approached after deformation due to pressure or detachment of a substrate. This behavior can be supported by subsequent crosslinking with heat or irradiation (electron beam, UV, HF) or the use of fillers.
Hereinafter the invention is illustrated by examples which are not meant to limit the invention thereto. On the contrary, the person skilled in the art can make suitable modifications on the basis of his technical knowledge without thereby going outside the scope of the invention.

Residuelessly redetachable contact-adhesive sheetlike structures