Mechanism and intervention of murine transfusion-related acute lung injury caused by anti-CD36 antibodies.

Anti-CD36 Abs have been suggested to induce transfusion-related acute lung injury (TRALI) upon blood transfusion, particularly in Asian populations. However, little is known about the pathological mechanism of anti-CD36 Ab-mediated TRALI, and potential therapies have not yet been identified. Here, we developed a murine model of anti-CD36 Ab-mediated TRALI to address these questions. Administration of mouse mAb against CD36 (mAb GZ1) or human anti-CD36 IgG, but not GZ1 F(ab')2 fragments, induced severe TRALI in Cd36+/+ male mice. Predepletion of recipient monocytes or complement, but not neutrophils or platelets, prevented the development of murine TRALI. Moreover, plasma C5a levels after TRALI induction by anti-CD36 Abs increased more than 3-fold, implying a critical role of complement C5 activation in the mechanism of Fc-dependent anti-CD36-mediated TRALI. Administration of GZ1 F(ab')2, antioxidant (N-acetyl cysteine, NAC), or C5 blocker (mAb BB5.1) before TRALI induction completely protected mice from anti-CD36-mediated TRALI. Although no significant amelioration in TRALI was observed when mice were injected with GZ1 F(ab')2 after TRALI induction, significant improvement was achieved when mice were treated postinduction with NAC or anti-C5. Importantly, anti-C5 treatment completely rescued mice from TRALI, suggesting the potential role of existing anti-C5 drugs in the treatment of patients with TRALI caused by anti-CD36.

[Study of the platelet GP specific antibodies and HLA antibodies expression in platelet transfusion refractoriness patients].

OBJECTIVE: To investigate the correlation between the platelet GP specific antibodies/HLA antibodies and platelet transfusion refractoriness (PTR). METHODS: Sixty-five patients with PTR were selected in this study and were genotyped for HLA-A and HLA-B as well as HPA systems by standard PCR-SSP assays. The platelet GP specific antibodies and HLA antibodies in serum and platelet elution were tested with a solid phase ELISA. RESULTS: The HLA-A/B antigens and the frequencies of HPA-1, 2, 4, 5, 6, 9, 15 antigens in PTR patients had no difference from those in healthy donors. The freguencies of HPA-3a and 3b were 0.65 and 0.35, respectively. There was statistical difference between the 65 PTR patients and the healthy donors in HPA-3 freguencies (P < 0.05). Twenty-four patients (36.9 %) only expressed HLA antibodies, and 14 (21.5%) expressed HLA and platelet GP specific antibodies. The highest expression of anti-HLA-A/B specific antibodies was -A*9(46.2 %)/-B*40(33.6%), respectively. In serum, GPIIb/IIIa was expressed (26.2%), followed by GPIa/IIa (21.5 %). In platelet elution, GPIIb/IIIa was expressed of 41.5% and GPIb/IX 41.5%. Pedigree study was carried out for 2 patients. The results showed that the platelet GP specific antibody/HLA antibody developed in PTR patients was highly related to the mismatch with the platelet antigen/HLA antigen in their parents. CONCLUSION: The expressions of the HLA and platelet GP specific antibodies are the most important reason in PTR, it's meaningful to explore the correlation between PTR and HLA and HLA-A/B antigen in guiding platelet transfusion.

[Expression of anti-platelet glycoprotein specific antibodies and anti-HLA antibodies in idiopathic thrombocytopenic purpura].

In order to investigate the expression of the anti-platelet glycoprotein specific antibodies and anti-HLA antibodies in idiopathic thrombocytopenic purpura (ITP), 45 patients with ITP were selected in this study. An easy PCR-SSP assay was used to detect single-nucleotide polymorphisms or deletion in HPA and HLA systems. The anti-platelet glycoprotein specific antibodies and anti-HLA antibodies in plasma or platelet eluate were tested with a solid phase ELISA. The results indicated that the anti-platelet glycoprotein specific antibodies were detected in plasma or platelet eluate of 45 patients, among which anti-GPIIb/IIIa/and anti-GpIb/IX were most common. Both the anti-platelet glycoprotein specific antibodies and anti-HLA antibodies were found in plasma of 11 patients. Pedigree investigation in 2 patients (case 37 and case 40) was carried out, the results showed that anti-platelet glycoprotein specific antibodies and anti-HLA antibodies detected in 2 patients closely related to incompatibility with platelet antigens and HLA antigens in parents. In conclusion, the results suggested that detection of the anti-platelet glycoprotein specific antibodies and anti-HLA antibodies in plasma or platelet eluate in combination with investigation of clinical manifestation of patients is important for diagnosis of idiopathic thrombocytopenic purpura.

[Biological appraisal of human bone marrow mesenchymal stem cells during ex-vivo expansion].

This study was aimed to investigate the characteristics of human bone marrow mesenchymal stem cells during ex-vivo expansion, MSCs were isolated from human bone marrow. At each passage, the characteristics of proliferation kinetics, osteogenic and adipogenic differentiation potential were analyzed, and cell morphology, surface markers were investigated as well. The karyotype analysis was done in different passage cells. The infection HIV, HCV, HBV and TP were detected by ELISA. Mycoplasma contamination in vitro was detected by PCR method. HLA-SBT was used to reanalyze the results of HLA antigens and alleles. STR genetic loci were detected by PCR in the MSC1, MSC2, MSC3 and MSC4. The results indicated that the proliferative ability and osteogenic potential decreased with the increase of passage number during culture expansion. The multiple differentiation potential of MSCs was maintained during their life span. Karyotype analysis showed that MSCs from 4 groups before passage 8 were normal. The expression of CD29, CD44, CD105, CD166 and CD73 were positive. The expression of CD14, CD34, CD45, CD80, CD86 were all negative. SBT was used to identify HLA-A, B, Cw, DRB1, DRPB1, DQ alleles in the MSC1, MSC2, MSC3, MSC4. The genetype of STR in the MSC1, MSC2, MSC3, MSC4 was different. MSC 3 was examined by TP-ELISA to confirm the infectious disease of TP. MSC2 was contaminated by mycoplasma at passage 5. It is concluded that culture expansion causes MSCs to gradually lose their stem cell properties. During ex-vivo expansion of MSCs, the osteogenic differentiation potential is decreased. MSCs before passage 8 can be a valuable subject for basic research and clinical application.