PMC8403076_01
The changes of beta2-MG, IL-6, VEGF, LDH. The series of beta2-MG, IL-6 are drawn on the primary axis, and the series of VEGF, LDH are drawn on the secondary axis.
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IL-6 were elevated. In addition, his serum level of vascular endothelial growth factor (VEGF) was normal (Figure 1). After the third course, VEGF level was declined, but IL-6 and LDH elevated again (Figure 1).
Flow cytometry of peripheral blood. (A-F) Before chemotherapy, the aberrant lymphocytes population expressed CD19, CD20, CD10, cKappa; (H-M) after three courses of chemotherapy, these became negative.
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The immunophenotyping by FC was consistent with the phenotype of CD5-, CD10+ monoclonal B lymphocytes (Figure 2).
The immunostaining of right cervical lymph node for cytokines. (A) (b-FGF;x200), positive; (B) (IL-1beta;x200), positive; (C) (IL-2;x200), positive; (D) (IL-6;x200), positive; (E) (IL-10,x200), positive; (F) (MUM-1x200), positive; (G) (PDGFx200), positive; (H) (TGF-betax200), positive; (I) (TNF-alphax200), positive; (J) (VEGFx200), positive; (K) HE, x40.
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The immunostaining of bone marrow before treatment. (A) (b-FGF;x200), positive; (B) (CD20;x100), positive; (C) (CD61x100), positive; (D) (IL-10;x200), positive; (E) HE 100x; (F) (IL-1beta;x200), positive; (G) (PDGFx200), positive; (H) (TGF-betax200), positive; (I) (VEGFx200), positive; (J) reticular fiber staining revealed diffuse fibrosis (MF-3) 100x; (K) (IL-2;x200), positive; (L) (IL-6;x200), positive; (M) (TNF-alphax200), positive.
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TNF-alpha, TGF-beta, PDGF, IL-1beta, IL-2, IL-6, IL-10. The reticular fiber staining was grade 3 (Figure 4).
PET/CT finding. (A-C) Before chemotherapy. (The largest lymph nodes was in the lower abdominal mesenteric region 68mm x 42mm, SUVmax =8.2). (D-F) After three courses of chemotherapy (The spleen reduced obviously, whose metabolism was normal (SUVmax=2.6)). (A) Widespread lymph node enlargement and hypermetabolism, splenomegaly, kidney hypermetabolism (blue arrows) and diffuse bone marrow hypermetabolism. Its inferior border approached the upper margin of pelvis (SUVmax= 2.9); (B) spinal hypermetabolism (blue arrow); (SUVmax =9.0); (C) iliac hypermetabolism (blue arrows); (D) no abnormal lymph nodes were found. The volume of spleen was smaller than before, and the metabolism of bone marrow, kidney metabolism were normal (red arrows); (E) normal spine metabolism, (SUVmax =3.3) (red arrow); (F) normal ilium metabolism (red arrows).
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IV: General hypermetabolic enlarged lymph nodes were be seen in both sides of diaphragm, especially in the abdomen; the metabolism of bone marrow and left kidney was elevated. In addition, the PET scan showed splenomegaly obviously without hypermetabolism, which were induced by MF probably (Figure 5). TNF-alpha were normal. The PET images showed improvement of the lymphadenopathy (Figure 5).
Fluorescence interphase in situ hybridization on lymph node biopsy. (A) IGH-BCL2 dual-colour dual-fusion probe displays two fused green and red signals, one isolated green and one isolated red signal in most of tumour nuclei with t(14;18) (q21;q32)/IGH-BCL2. Arrow indicates positive nuclei. (B) BCL2 dual-colour break-apart (BAP) probe shows one co-localised green and red signal, one isolated green and one isolated red signal in most of tumour nuclei; (C) BCL6 dual-colour break-apart (BAP) probe shows two co-localised green and red signal; (D) MYC dual-colour break-apart (BAP) probe shows one co-localised green and red signal.
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Fluorescence in situ hybridization (FISH) analysis performed on paraffin embedded tissue sections from the right cervical lymph node were positive for the BCL2 (18q21) split signal and IGH/BCL2 fusion signal, and negative for MYC rearrangement and BCL6 (3q27) split signals (Figure 6).
The immunostaining of bone marrow after one course of chemotherapy. (A) (b-FGF;x200), positive; (B) (CD19;x200), negative; (C) (CD117x200), negative; (D) (IL-1beta;x200), positive; (E) (IL-2;x200), positive; (F) (IL-6;x200), positive; (G) (IL-10;x200), positive; (H) (PDGFx200), positive; (I) (TGF-betax200), positive; (J) (TNF-alphax200), positive; (K) (VEGFx100), positive; (L) reticular fiber staining x200, MF-1.
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The immunostaining of bone marrow showed the tumor cells were negative for CD19, CD117, b-FGF, TNF-alpha, TGF-beta, PDGF, IL-1beta, IL-2, IL-6, IL-10, except for VEGF after two courses (Figures 7 and 8). The reticular fiber staining was grade 1 (Figure 7).
The immunostaining of bone marrow after two courses of chemotherapy. (A) (b-FGF;x200), negative; (B) (CD19;x200), negative; (C) (CD117;x200), negative; (D) (IL-1beta;x200), negative; (E) (IL-2;x100), negative; (F) (IL-6;x200), negative; (G) (IL-10;x200), negative; (H) (PDGFx200), negative; (I) (TGF-betax200), negative; (J) (TNF-alphax200), negative; (K) (VEGFx100), positive; (L) reticular fiber staining 100x, MF 0-1.
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The immunostaining of bone marrow showed the tumor cells were negative for CD19, CD117, b-FGF, TNF-alpha, TGF-beta, PDGF, IL-1beta, IL-2, IL-6, IL-10, except for VEGF after two courses (Figures 7 and 8).