Background: Acute graft?versus?host disease (aGVHD) is a common and severe complication of allogeneic hematopoietic stem cell transplantation (allo?HSCT). Some studies have found that the presence of certain specific human leukocyte antigen (HLA) loci could affect the occurrence of aGVHD. Meanwhile, the impact of HLA haplotypes on aGVHD has been rarely studied. This study aimed to investigate the effects of HLA loci and haplotypes on intestinal aGVHD. Methods: Totally, 345 consecutive patients undergoing first HLA?matched sibling peripheral blood stem cell transplantation (PBSCT) from February 2004 to June 2013 at Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, were enrolled in this study. HLA loci and haplotypes of recipients with frequency over 5% were searched and their effects on intestinal aGVHD were investigated. Other important factors including donor age, recipient age, donor?recipient sex combinations, and conditioning regimens were also evaluated using logistic regression. Pure upper gastrointestinal tract aGVHD without diarrhea was excluded because the histological proof was unavailable. The follow?up end?point was 6 months after HSCT. Results:The cumulative incidence of intestinal aGVHD was 19.4%,with 18.0%of the patients classified as classic aGVHD and 1.4% aspersistent, recurrent,or late aGVHD.Multivariate analysis showed that HLA?A31 locus(odds ratio[OR]2.893,95% confidence interval[CI][1.054,7.935], P = 0.039), HLAB40?DR15 (OR 3.133, 95% CI [1.250, 7.857], P = 0.015), and HLAB46?DR9 haplotypes (OR 2.580, 95% CI [1.070, 6.220], P = 0.035), female donor for male recipient (OR 2.434, 95% CI [1.319, 4.493], P = 0.004) were risk factors for intestinal aGVHD. Conclusion: The presence of certain HLA loci and haplotypes may influence the occurrence of intestinal aGVHD in PBSCT with HLA?identical sibling donors.

Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Transplantation Conditioning ; Transplantation, Homologous

OBJECTIVETo investigate the molecular genetics basis for HLA novel allele HLA-DRB1*1212 in Chinese population.

METHODSGenomic DNA was extracted from whole blood by salting-out method. HLA-DRB1 gene exon 2 was amplified by PCR with group-specific primers from genomic DNA. PCR products were cut back from agarose gels and purified to sequence directly. The polymerase chain reaction-sequence specific oligonucleotide probes (PCR-SSO) was performed to confirm the mutations which were detected by sequencing in this study.

RESULTSThe sequencing results showed HLA-DRB1 alleles of the proband as DRB1*090102 and the novel allele. The sequences of the novel allele have been submitted to GenBank (AY899825). Through BLAST analysis, the novel allele was found to be different from DRB1*120101 at position 199A-->C in exon 2, that results in an amino acid change from Ile to Leu at codon 67.

CONCLUSIONThis allele is a novel and has been officially named as DRB1*1212 by the WHO Nomenclature Committee.

Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; ethnology ; DNA ; analysis ; Ethnic Groups ; Female ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Analysis

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; drug effects ; Ethanol ; adverse effects ; Hepatocytes ; cytology ; Liver Regeneration ; Mesenchymal Stromal Cells ; cytology ; Rats ; Rats, Sprague-Dawley

The study was purposed to investigate the effect of alloreactive natural killer (alloNK) cells on immune reconstitution in murine haploidentical bone marrow transplantation (BMT). The murine model of haploidentical BMT was established by using (C57BL/6×BALB/c)BCF(1)(H-2(d/b)) mouse as the donor, and BALB/c (H-2(d)) mouse as the recipient. Recipient mice were divided into BMT group, non-allo-reactive NK (non-alloNK) cell group and alloNK cell group according to different transfusion. The effect of adding alloNK cells to transfusion was assessed by thymus pathology, the proportion of spleen NK cells, the spleen cell proliferation, the IFN-γ and IL-4 concentrations product at 24 and 48 h of recipient spleen cell culture supernatant at 2 months after BMT. The results showed that there were no obvious difference in thymus tissue among 3 groups under the optical microscope. The proportion of recipient spleen NK cells in non-alloNK group was significantly lower than that in BMT group (P < 0.05). There was no significant difference in proliferation of the recipient spleen cells among 3 groups at 2 months after BMT. The IFN-γ concentration product at 24 and 48 h of recipient spleen cell culture supernatant in alloNK group was significantly lower than that in other 2 groups at 2 months after BMT (P < 0.05). The IL-4 concentration in each group was not significantly different (P > 0.05). It is concluded that alloNK cells do not damage the thymus structure and may induce Th2 immune response in murine haploidentical BMT.
Animals ; Bone Marrow Transplantation ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Killer Cells, Natural ; immunology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Spleen ; cytology ; Thymus Gland ; pathology ; Transplantation, Homologous

The aim of this study was aimed to investigate the molecular genetic basis for a novel HLA allele, HLA-B*4061, in Chinese population. DNA was extracted from whole blood by salting-out method. The HLA-B exons 1 - 8 of the proband was amplified and the amplified product was cloned using TOPO TA cloning sequencing kit to split the two alleles apart. Both strands of exons 2, 3 and 4 of chosen colonies were sequencing. The PCR-SSP was performed to confirm the mutations detected by sequencing. The sequencing results showed HLA-B alleles of the proband as B*4601 and the novel allele. The sequences of the novel allele have been submitted to GenBank (DQ089628, DQ089629, DQ089630). After HLA blast analysis, the novel allele showed a single nucleotide mismatch with B*400101 in exon 2 at position 272 C-->A, as the results, changing amino acid from Ser to Tyr at codon 67. It is concluded that this allele is a novel one and has been officially named B*4061 by the WHO Nomenclature Committee.
Alleles ; Amino Acid Substitution ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; HLA-B Antigens ; genetics ; immunology ; Histocompatibility Testing ; Humans ; Molecular Sequence Data ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid

To explore the antitumor activities of KRN7000 in a NS-1 myeloma-bearing mice and the underlying mechanisms, cytotoxic activities of the spleen cells treated with KRN7000 in vivo and in vitro were examined with Yac-1 (a NK-sensitive cell line) and NS-1 (a NK-insensitive cell line) as the targeted cells. The life span of NS-1 myeloma-bearing mice was observed after treating with KRN7000 and combined with cyclophosphamide (CTX). Furthermore, toxicities of KRN7000 administration on liver and kidney were evaluated. The results showed that KRN7000 enhanced NK cytotoxic activities of spleen cells. Around 20% of NS-1-bearing mice survived after KRN7000 administration and the survival rate reached up to 80% of NS-1-bearing mice when KRN7000 was used in combination with CTX at a dose of 100 mg/kg (P < 0.005). KRN7000, at a dose of 100 micro g/kg, had no toxic effects on liver and kidney. These findings suggest that KRN7000 might be a promosing agent in tumor immunotherapy.

OBJECTIVETo investigate the risk factors of intestinal acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSThe clinical data of 534 cases of 533 patients undergoing allo-HSCT during Jan 2004 and Sep 2012 were retrospectively analyzed. The effects of donor-recipient HLA mismatching, recipient age, donor age, donor-recipient sex combination, donor-recipient relationship, HSC source, conditioning regimen with or without total body irradiation (TBI) and HLA loci on intestinal aGVHD with different severity were analyzed by Logistic regression.

RESULTSIntestinal aGVHD occurred in 123(23.0%) cases, with 86(16.1%) cases of stage 1 intestinal aGVHD(16.1%) and 37(6.9%) cases of stage 2 to 4 intestinal aGVHD. Multivariate analysis showed that donor-recipient HLA mismatching (OR=2.519, P=0.002), increasing donor age (OR=1.034, P=0.003), female donor for male recipient (OR=1.855, P=0.007) were risk factors for intestinal aGVHD, HLA-B38 (OR=0.256, P=0.032) was its protective factor. Donor-recipient HLA mismatching (OR=2.799, P=0.011), increasing donor age (OR=1.045, P=0.012), HLA-A1 (OR=4.157, P=0.002), A30 (OR=3.143, P=0.005) were risk factors for stage 2 to 4 intestinal aGVHD.

CONCLUSIONOccurrence of intestinal aGVHD and its severity are associated with donor-recipient HLA mismatching, donor age, donor-recipient sex relationships and some HLA loci.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Graft vs Host Disease ; epidemiology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Intestinal Diseases ; epidemiology ; Male ; Middle Aged ; Retrospective Studies ; Risk Factors ; Tissue Donors ; Transplantation, Homologous ; adverse effects ; Young Adult