PMC8369751_01
Clinical history (Timeline). R, Rituximab; CHOP: C, cyclophosphamide; H, hydroxyadriamycine; O, oncovin ; P, prednisone; CT, computerized tomography; ATB, antibiotherapy.
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Lungs were not involved at this time (Supplementary Figure S1). Timeline of his medical history is resumed in Figure 1.
Clinical evolution and histological analysis of the patient. (A) 18F-FDG-TEP-CT-scan revealing an interstitial lung disease with nodular lesions (left). CT-scan normalization after 3 months (right). (B) Histological analysis showing a peri-bronchial granulomatous inflammation with ulceration (upper left), a perivascular granuloma (upper right), giant multinucleated cells (lower left), and BAAR with Ziehl-Neelsen staining (lower right).
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Association Classification class II-IV). An interstitial lung disease with nodular lesions was diagnosed on CT-scan with a decline of alveolar exchanges on breath tests (Figure 2A). A lung biopsy was performed that finally revealed invasive infection by M. bovis BCG, suspected by Ziehl-Neelsen coloration and confirmed by PCR and culture (Figure 2B). Table S1). This strategy finally allowed an improvement of the patient's condition with CT-scan and TEP-scan complete normalization after 3 and 6 months, respectively (Figure 2A).
Absence of IFNgamma or other autoantibodies that may directly hinder/block IFNgamma signaling and normal response to IFNgamma or BCG stimulation. STAT1 phosphorylation (Y701) of HD monocytes after stimulation for 15 min (A) with 25% (v/v) allogenic HD or patient's serum (from different time points: P1, P2, and P3) and (B) with IFNgamma previously mixed with 25% (v/v) allogenic HD or patient's serum (from different time points: P1, P2, and P3). (C) STAT1 phosphorylation (Y701) of HD monocytes after IFNgamma stimulation for 15 min. Cells were preincubated 30 min at room temperature in the presence of 25% (v/v) HD or patient's serum (from different time points: P1, P2, and P3) and washed before IFNgamma stimulation. (D) STAT1 phosphorylation (Y701) of monocytes from the patient after IFNgamma stimulation for 15 min. (E) Frequency of IFNgamma+ and/or TNFalpha+ CD4+ T cells from the patient after no stimulation or stimulation with heat inactivated BCG for 48 h. (F) IFNgamma concentration in culture supernatant after no stimulation, IL12, heat-inactivated BCG, or PMA/IONO stimulation of PBMC from the patient and one HD for 48 h. BCG: bacillus Calmette-Guerin; HD, healthy donor; IFNgamma, interferon gamma; NS, non-stimulated; P1, patient's serum before BCGosis diagnosis and IFNgamma therapy (2018-08); P2, patient's serum at the time of BCGosis diagnosis and during IFNgamma therapy (2018-11); P3, patient's serum after BCGosis diagnosis and IFNgamma therapy (2019-02).
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These results suggested for the absence of anti-IFNgamma autoantibodies or any other autoantibodies that may directly hinder/block IFNgamma signaling (Figures 3A-C). STAT1 phosphorylation, after IFNgamma stimulation of monocytes from the patient, was preserved (Figure 3D). CD4+ T cells from the patient responded to BCG (Figure 3E) and showed no apparent defect of IFNgamma production after BCG +- IL12 stimulation (Figure 3F).
Maximum likelihood phylogenetic tree representing the relationship between the clinical M. bovis BCG strain (M. bovis BCG_1811074784) and reference M. bovis BCG strains. Bootstrap values of each branch are indicated.
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The results showed that the patient's strain (M. bovis BCG_ 1811074784, Figure 4) was most closely related to the M. bovis BCG Tice strain.