Humans ; Neutrophils ; Receptors, Cell Surface ; GPI-Linked Proteins ; HIV Infections ; Isoantigens

Graft-versus-host disease (GVHD), a life-threatening complication after bone marrow transplantation (BMT), is induced by activation of alloreactive donor T cells. Our previous study demonstrated that transplantation of myeloid differentiation factor 88 (MyD88)-deficient knockout (KO) bone marrow (BM) resulted in aggravation of GVHD. Here, to understand the cellular mechanism, we performed longitudinal in vivo imaging and flow cytometric analyses followed by transcriptome and functional examination of donor MyD88-KO BM progenies in GVHD hosts, using a major histocompatibility complex-matched but minor histocompatibility antigen-mismatched C57BL/6→BALB.B model. In GVHD hosts with MyD88-KO BMT, donor BM-derived CD11b+Gr-1+ cells were found to undergo cell death, a fate significantly different from the explosive expansion shown by the wild type (WT) counterparts, and also from the moderate expansion of the WT or MyD88-KO BM-derived cells in non-GVHD hosts. It was also revealed that MyD88-KO CD11b+Gr-1+ cells preferred differentiation into CD11c+ dendritic cells (DCs) to expansion as myeloid-derived suppressor cells in GVHD hosts or in high inflammatory in vitro conditions. These CD11c+ DCs comprised the majority of MyD88-KO CD11b+Gr-1+ apoptotic cells in GVHD hosts. Their ability to cross-present alloantigens of host origin contributed to the enhancement of T cell alloreactivity, causing GVHD aggravation and eventually death through the killing function of activated T cells. These results provide insights into the roles of MyD88 in myelopoiesis of donor BM and the protective effects in GVHD hosts, helpful information for development of a strategy to control GVHD.
Bone Marrow Cells* ; Bone Marrow Transplantation ; Bone Marrow* ; Cell Death ; Dendritic Cells* ; Graft vs Host Disease ; Histocompatibility ; Homicide ; Humans ; In Vitro Techniques ; Isoantigens ; Myeloid Differentiation Factor 88 ; Myelopoiesis ; T-Lymphocytes ; Tissue Donors* ; Transcriptome

B cells play a role in graft rejection via several mechanisms. Specifically, B cells produce high-affinity antibodies to alloantigens including allogeneic major histocompatibility complex (MHC) with the help of follicular helper T cells. B cells also function as antigen-presenting cells for alloreactive T cells, resulting in the activation of alloreactive T cells. Conversely, the frequency of regulatory B cells increases under inflammatory conditions and suppresses the rejection process. Here, the differential roles of the major B cell subpopulations (B-1, follicular B, marginal zone B, and regulatory B cells) involved in transplantation rejection are discussed together with their interaction with T cells.
Antibodies ; Antibody Diversity ; Antigen-Presenting Cells ; B-Lymphocytes* ; B-Lymphocytes, Regulatory ; Graft Rejection* ; Isoantigens ; Major Histocompatibility Complex ; T-Lymphocytes ; T-Lymphocytes, Helper-Inducer

Inbred mice, a systematically developed homogeneous animal, have been developed to maintain experimental reproducibility and to minimize experimental variables in animal-based studies. In particular, C57BL/6 mice are frequently used in experiments into immunology and antitumor activity experiments. This study was compared the immunological characteristics of C57BL/6NKorl, a Korean developed experimental animal resource, with those of two other C57BL/6N substrains. Mouse body, thymus, and spleen weights in C57BL/6NKorl were not significantly different from those of the other two C57BL/6N substrains. Among the three substrains, there was no difference in the distribution of T and B cells, which are lymphocytes involved in adaptive immunity, and no difference in NK cells related to innate immunity. Results for macrophages and granulocytes, which have roles in innate immunity, were similar in all three substrains. In order to investigate the expressions of major histocompatibility complex (MHC) molecules and allogenic antigens, splenocytes were separated from obtained spleen and analyzed by using flow cytometry. MHC class I and II molecules, which are important during self/non-self-discrimination, were similar in the three substrains. In addition, expression of alloantigen involved in allografts showed similar results in the three substrain. Thus, the results of this study provide strong evidence that C57BL/6NKorl is immunologically similar to two other C57BL/6N substrains.
Adaptive Immunity ; Allergy and Immunology ; Allografts ; Animals ; B-Lymphocytes ; Flow Cytometry ; Granulocytes ; Immunity, Innate ; Isoantigens ; Killer Cells, Natural ; Lymphocytes ; Macrophages ; Major Histocompatibility Complex ; Mice* ; Spleen ; Thymus Gland ; Weights and Measures

BACKGROUND: Alloantibodies against human neutrophil alloantigen (HNA)-3a are associated with severe and fatal transfusion related acute lung injury (TRALI). HNA-3 genotyping and HNA-3a antibody (Ab) identification are essential to diagnosis and prevention of TRALI caused by HNA-3a Ab. However there had been no laboratory for HNA-3a Ab identification in Korea. The aims of this study were to establish the HNA-3a Ab test in Korea and to estimate the incidence of HNA-3a alloimmunization among pregnant Korean women. METHODS: HNA-3a homozygotes and HNA-3b homozygotes were identified by HNA-3 genotyping. Three HNA-3a homozygotes and three HNA-3b homozygotes are included in the granulocytes panel, which consisted of 10 donors for granulocytes. Sera from 650 pregnant Korean women were tested for granulocyte Ab using a mixed passive hemagglutination assay (MPHA). When a HNA-3a Ab was detected, the woman's HNA-3 was typed to support her HNA-3a alloimmunization. RESULTS: MPHA showed positive reactions in the sera from 26 women (4.0%, 26/650). HLA Abs were detected in 18 women (2.8%, 18/650), among whom HNA Abs were identified simultaneously in 7 women. Granulocyte Abs were detected in sera from 15 women (2.3%, 15/650). The incidence of HNA-3a, HNA-1b, HNA-1a, HNA-2a, and unidentified HNA Abs among pregnant Korean women was 0.77% (5/650), 0.77% (5/650), 0.62% (4/650), 0.15 (1/650), and 0.31% (2/650), respectively. CONCLUSION: In this study, we established the HNA-3a Ab test using MPHA for diagnosis and prevention of TRALI caused by HNA-3a Ab. The incidence of HNA-3a Ab in pregnant Korean women was 0.77% (5/650).
Acute Lung Injury ; Diagnosis ; Female ; Granulocytes ; Hemagglutination ; Homozygote ; Humans* ; Incidence ; Isoantibodies ; Isoantigens ; Korea ; Neutrophils* ; Tissue Donors

Neutrophils play an essential role in the innate immune response to infection. Neutrophils migrate from the vasculature into the tissue in response to infection. Recently, a neutrophil cell surface receptor, CD177, was shown to help mediate neutrophil migration across the endothelium through interactions with PECAM1. We examined a publicly available gene array dataset of CD177 expression from human neutrophils following pulmonary endotoxin instillation. Among all 22,214 genes examined, CD177 mRNA was the most upregulated following endotoxin exposure. The high level of CD177 expression is also maintained in airspace neutrophils, suggesting a potential involvement of CD177 in neutrophil infiltration under infectious diseases. To determine the role of CD177 in neutrophils in vivo, we constructed a CD177-genetic knockout mouse model. The mice with homozygous deletion of CD177 have no discernible phenotype and no significant change in immune cells, other than decreased neutrophil counts in peripheral blood. We examined the role of CD177 in neutrophil accumulation using a skin infection model with Staphylococcus aureus. CD177 deletion reduced neutrophil counts in inflammatory skin caused by S. aureus. Mechanistically we found that CD177 deletion in mouse neutrophils has no significant impact in CXCL1/KC- or fMLP-induced migration, but led to significant cell death. Herein we established a novel genetic mouse model to study the role of CD177 and found that CD177 plays an important role in neutrophils.
Animals ; Disease Models, Animal ; GPI-Linked Proteins ; genetics ; Gene Expression Regulation ; Genetic Therapy ; Humans ; Immunity, Innate ; genetics ; Inflammation ; genetics ; microbiology ; pathology ; Isoantigens ; genetics ; Mice ; Mice, Knockout ; Neutrophils ; metabolism ; pathology ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Receptors, Cell Surface ; genetics ; Staphylococcus aureus ; pathogenicity ; Transcriptional Activation

OBJECTIVETo explore the value of HLA-DRB1 gene in predicting the outcome of unexplained recurrent spontaneous abortion (URSA) treated with paternal lymphocyte alloimmunization therapy (PLAT) in Henan Hans.

METHODSThree hundred URSA patients were recruited. Following PLAT treatment, they were divided into two groups according to the outcome of pregnancy. Polymerase chain reaction sequence specific primer (PCR-SSP) were conducted to analyze the HLA-DRB1 gene.

RESULTSFor those who have received PLAT treatment, the frequency of HLA-DRB1*11 was significantly lower in successfully treated cases than those with abortion (0.052 vs. 0.110, P < 0.05, OR=0448), whilst the frequency of HLA-DRB1*15 was significantly greater in the former (0.207 vs. 0.100, P < 0.05, OR=2.352).

CONCLUSIONFor patients who have received PLAT treatment, those with HLA-DRB1*15 are more likely to conceive that those with HLA-DRB1*11.

Abortion, Spontaneous ; ethnology ; genetics ; immunology ; therapy ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; Female ; Genetic Predisposition to Disease ; ethnology ; HLA-DRB1 Chains ; genetics ; Humans ; Immunotherapy ; Isoantigens ; immunology ; Lymphocytes ; immunology ; Male ; Pregnancy ; Treatment Outcome

Neonatal alloimmune thrombocytopenia (NAIT) occurs when maternal alloantibodies react to antigens expressed on fetal platelets, which is mainly platelet-specific alloantigen or HLA, resulting in their immune destruction. Here, we described a patient who suffered from NAIT caused by anti-HLA-A2 antibody. Sera from the mother and the newborn were screened for human platelet antigen-specific antibodies and HLA antibodies by ELISA, and HLA antibodies were detected in both of them. The antibody specificity was identified as anti-HLA-A2 by Luminex single antigen bead assay. HLA typing results showed that patient's father descended HLA-A2 antigen on the patient and the mother was HLA-A2 negative. It is most conceivable that anti-HLA-A2 alloantibody in the mother's sera crossed the placenta and subsequently caused NAIT in the case presented. The patient received platelet concentrates, oral steroid and intravenous globulin and platelet count increased to 120x10(9)/L on the 90th day of life. The Luminex single antigen bead assay used in this case is highly sensitive and specific assay to determine antibody specificity and it is faster and more convenient for routine use in clinical laboratory so that this assay could be useful to diagnose NAIT caused by HLA antibodies and treat such NAIT patients with HLA matched platelet transfusion.
Antibodies ; Antibody Specificity ; Blood Platelets ; Enzyme-Linked Immunosorbent Assay ; Fathers ; Histocompatibility Testing ; HLA-A2 Antigen ; Humans ; Infant, Newborn ; Isoantibodies ; Isoantigens ; Mothers ; Placenta ; Platelet Count ; Platelet Transfusion ; Thrombocytopenia, Neonatal Alloimmune

No abstract available.
Animals ; Antigen-Presenting Cells/immunology ; Graft Rejection/immunology/*prevention & control ; Graft Survival ; *Histocompatibility ; Humans ; Immunosuppression/*methods ; Intercellular Adhesion Molecule-1/immunology ; Isoantigens/*immunology ; Organ Transplantation/*adverse effects ; *Transplantation Tolerance

PURPOSE: To treat burn and burn scar contracture, many types of skin grafts have been developed. Skin allograft is a recently introduced option for reconstruction of skin defects. The science of skin allotransplanation is rooted in progressive thinking by surgeons, fueled by innovative solutions, and aided by understanding the immunology of tolerance and rejection. METHODS: This study assesses the effect of dendritic cell pretreatment in induction of survival increase in a rat skin allograft model. Recipient derived dendritic cells were harvested from rat whole blood and cultured with GM-CSF and IL-4 for 2 weeks. Then donor-specific alloantigen pulsed dendritic cells were reinjected into subdermal tissue before skin graft. Rat left dorsal skin allografts were transplanted in 3 subgroups. Groups: I) untreated, II) FK-506 (2 mg/kg), III) dendritic cell pretreatment and FK-506. Graft appearance and histologic analysis were assessed postoperatively. RESULTS: The group III showed longest graft survival rate than other groups. CONCLUSION: Donor antigen pulsed host dendritic cell combined with short-term immunosuppression prolong skin allograft survival and has potential therapeutic application for induction of tolerance.
Animals ; Burns ; Cicatrix ; Contracture ; Dendritic Cells ; Graft Survival ; Granulocyte-Macrophage Colony-Stimulating Factor ; Humans ; Immunosuppression ; Interleukin-4 ; Isoantigens ; Rats ; Skin ; Tacrolimus ; Thinking ; Tissue Donors ; Transplantation, Homologous ; Transplants