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The five closely linked CD1A-E genes encode the human CD1 family of proteins. Few studies of the allele frequencies of these genes in African populations have been published so far. This study aimed to genotype CD1A and CD1E variants and to compare their frequencies in Sub-Saharan Africans from Gabon and Ivory Coast, and Non-Africans from Syria and France. A restriction analysis of DNA fragments generated by PCR was performed to detect CD1A and CD1E alleles in 105 subjects from Gabon, 169 subjects from Ivory Coast, 107 subjects from Syria and 181 subjects from France. The frequencies of the CD1E*02 allele were high among Sub-Saharan Africans (87%) and low in West Asians (44%) and Europeans (36%), whereas the contrary was obtained for the CD1E*01 allele (7%, 55% and 64% respectively). Frequencies of CD1A alleles were similar between all groups, the CD1A*02 allele was most prevalent (91%). The high frequency of the CD1E*02 allele in Sub-Saharan Africans suggest that future work should investigate the relationship between CD1 polymorphism and infectious diseases.

Background Alloimmunization or the development of alloantibodies to Red Blood Cell (RBC) antigens is considered one of the major complications after RBC transfusions in patients with sickle cell disease (SCD) and can lead to both acute and delayed hemolytic reactions. It has been suggested that polymorphisms in HLA genes, may play a role in alloimmunization. We conducted a retrospective study analyzing the influence of HLA-DRB1 and DQB1 genetic diversity on RBC-alloimmunization. Study design Two-hundred four multi-transfused SCD patients with and without RBC-alloimmunization were typed at low/medium resolution by PCR-SSO, using IMGT–HLA Database. HLA-DRB1 and DQB1 allele frequencies were analyzed using logistic regression models, and global p -value was calculated using multiple logistic regression. Results While only trends towards associations between HLA-DR diversity and alloimmunization were observed, analysis of HLA-DQ showed that HLA-DQ2 ( p = 0.02), -DQ3 ( p = 0.02) and -DQ5 ( p = 0.01) alleles were significantly higher in non-alloimmunized patients, likely behaving as protective alleles. In addition, multiple logistic regression analysis showed both HLA-DQ2/6 ( p = 0.01) and HLA-DQ5/5 ( p = 0.03) combinations constitute additional predictor of protective status. Conclusion Our data suggest that particular HLA-DQ alleles influence the clinical course of RBC transfusion in patients with SCD, which could pave the way towards predictive strategies.

A soluble isoform of MHC class I chain-related molecule A (soluble MICA), generated by proteolytic shedding from the membrane-bound MICA of various tumor cells, has been shown to downregulate both the expression of natural killer group 2-member D receptor and the cytotoxic function of effectors cells and was postulated as a mechanism for tumor immune evasion. Its effect on the expression of cytokines by the effector cells remained unexplored. Here we demonstrate that the sMICA molecules upregulate interferon gamma expression by interleukin-12/interleukin-18-activated CD3(-)CD56(+) natural killer cells, witnessing the pro-inflammatory effect of soluble MICA. Overall, these data are in line with our previous observations that the raised serum levels of soluble MICA, following allogeneic hematopoietic stem cell transplantation, confer susceptibility to and the presence of pre-transplantation anti-MICA antibodies in the patient's serum confer protection against chronic graft versus host disease.

Aim Strong or weak NK/T cell activation by NKG2D-MICA-129 met or val allele respectively can lead to different allograft outcomes by influencing the degree of NK cell activation. The aim was to understand the role of MICA-129 dimorphism in allograft rejection in live donor renal transplantation. Methods A total of 176 live donor renal transplant patients were genotyped for MICA-129 dimorphism using TaqMan technique and were followed up for a minimum period of 1.5 years post-transplant. Serum samples of all recipients were evaluated at the pre and post-transplant periods, 1–2 week, 3 months, 6 months and at time of rejection (if applicable) for the presence of HLA specific antibodies using Luminex technology. The biopsy proven rejection episodes were recorded and correlated with MICA-129 dimorphism for the presence of met or val alleles and the respective genotypes met/met, met/val and val/val. Results MICA-129 met allelic as well as met/met genotypic frequencies dominated in recipients experiencing rejection episodes, particularly those with antibody mediated rejection (AMR) as compared with the no rejection episode group (NRE) (61% vs 32%; p > 0.0006 and 43% vs 13%; p > 0.01 respectively). Five of the six met/met AMR cases developed denovo high titer (Mean MFI > 5000) HLA-DSA for HLA class II molecules, 3–6 months post transplantation. This is in contrast to the met/val and val/val groups who maintained low titer antibodies till the time of biopsy. Presence of met allele also correlated with serum creatinine levels and C4d positivity and the data revealed an allele dose dependent increase in intensity towards AMR progression. Conclusions These results point towards an important role of MICA-129 dimorphism in renal allograft rejection and suggest that recipients with MICA-129 met allele may warrant special post-transplant intervention/management.

Severe bacterial infections are the major causes of morbidity and mortality in sickle cell anemia (SCA) but are poorly explained. The distribution of a bi-allelic polymorphism (Arg107Gly) of human leukocyte antigen-E (HLA-E) locus was investigated in 144 SCA patients, most of whom originated from from sub-Saharan Africa. Among them, 73 presented with at least one severe bacterial infection, whereas 71 did not. The HLA-E*0101/E*0101 genotype was more frequent among the group with infections than their counterparts (47% vs 21%; p corrected = 0.003). This genetic association is of relevance, given the emerging evidence for the involvement of HLA-E molecules in host response to pathogens.

More and more cells and tissues of human origin are considered as unique medical bio-resources and are instrumental in the ongoing development of regenerative medicine. Although a better understanding of the genetic complexity of the major histocompatibility complex has contributed to develop allogeneic hematopoietic stem cell transplantation, the autologous setting was viewed early on only as a means to overcome myelodepression. The contribution of the immune system for adoptive immunotherapy was recognized in the 1990s, when donor lymphocyte infusion demonstrated an antileukemic effect of the transfused T cells. This led to the foundation and development of adoptive immunotherapy in the treatment and prevention of infectious diseases and cancer. The recognition that the immune system is undergoing a progressive decline and deterioration with age introduces a new challenge. Immunosenescence results in a well-documented increase in incidence and severity of infections, impaired responses to vaccines, and development of cancer. We propose and argue that autologous white blood cells collected and cryopreserved at a young age will represent a valuable bio-resource for the restoration of immunity and the successful development of adoptive immunotherapies in treating infections and cancer, and will pave the way to anticipatory medicine.

Human CD1 group I molecules CD1a, b, and c are expressed on antigen-presenting cells, notably dendritic cells, and implicated in glycolipids presentation to T lymphocytes. Expression of CD1 on monocytes is a hallmark of their activation. Because monocyte activation has been reported during steady state disease in sickle cell anemia (SCA) patients, we have analyzed CD1 expression on monocytes from 45 SCA patients originating from Africa and 27 healthy control subjects. CD1 expression was detected on monocytes in the majority of SCA patients (75%), whereas it was not observed in the vast majority of the control group (70.4%). CD1b and CD1c were highly expressed in Sβ thalassemia patients and CD1a expression was predominant in SDPunjab patients. This expression of the CD1 molecules is correlated with an increased expression of the major histocompatibility complex class II invariant chain (CD74). Finally, we have observed that the majority of SCA patients (68%) express only two or one CD1 isoforms. This study demonstrates the particular phenotype of SCA monocytes intermediate between normal resting and activated monocytes, a phenotype that could have consequences on regulation of the infection outcome.

Human leukocyte antigen (HLA) class II heterodimers have a well defined role in peptide presentation to helper T lymphocytes. Moreover, engagement of HLA class II molecules leads to signal transduction in the antigen presenting cell. Signaling via HLA-DR increases CD95 mediated hypoploidy in B-cell blasts. Given the importance of CD95 for the homeostasis of lymphocyte populations, we examined the impact of the HLA-DR signal on the most proximal events in the CD95 apoptotic pathway. CD95 activation recruits the adapter molecule Fas-associated death domain protein (FADD) and thereby provides a scaffold for procaspase-8 activation. The HLA-DR signal increased both recruitment of FADD and activation of caspases-3 and -8 via CD95 activation. Sensitization was tightly controlled because neither FADD recruitment to CD95 nor caspase activation was induced via HLA-DR alone. In contrast, the HLA-DR signal and the CD95 signal both led to decreased mitochondrial membrane potential. Taken together, these data indicate that ligand interaction with HLA class II molecules preactivates the antigen presenting cell for death in the event of a subsequent interaction with the CD95 ligand. This would ensure termination of a specific immune response, particularly in cells with limited sensitivity to CD95 mediated apoptosis alone.

Tumor necrosis factor alpha (TNFalpha) is a pleiotropic cytokine involved in inflammatory responses which can trigger both cell apoptosis and cell activation. In antigen presenting cells (APC), TNFalpha increased antigen presentation, notably by up-regulation of HLA class II expression. In addition to their role in antigen presentation, HLA-DR molecules transduce intracellular signals which lead to cytokine up-regulation or cell death. We have previously observed that the susceptibility of APC to HLA-DR mediated apoptosis increase throughout their maturation. We therefore investigated the relationship between TNFalpha production and susceptibility to HLA-DR-mediated apoptosis of different APC. The hematopoietic progenitor cell line (KG1), monocytic cell line (THP-1), monocyte-derived dendritic cell (DC), and B-lymphoid cell line (Raji) have been studied. We report that apoptosis susceptibility and spontaneous TNFalpha release are correlated in these different cells. However, while autocrine TNFalpha production was critical for DC maturation, upregulation of TNFalpha release after HLA-DR crosslinking was not observed and neutralization of endogenous TNFalpha did not modify HLA-DR-mediated apoptosis. These data reveal that HLA-DR mediated apoptosis susceptibility and spontaneous TNFalpha release are regulated in a parallel manner and that while TNFalpha may induce maturation of APC to an "apoptosis sensitive" stage, there is no direct role for TNFalpha in HLA-DR-mediated apoptosis of APC.

A novel HLA-B∗39 variant, found in an African patient with sickle cell anemia undergoing bone marrow transplantation is described. Initially suspected by inconsistent serological typing (B-blank, Bw6), then recognized by PCR-SSP, and finally characterized by nucleotide sequencing, this novel allele is designated HLA-B∗3916. It differs from HLA-B∗3910 by a point mutation (G to C) at position 17 of exon 3 causing glutamine to histidine change at codon 96 of α 2 domain, a conserved position among HLA class I alleles. cDNA sequence analysis further revealed the presence of both normally and abnormally spliced mRNA species in established cell lines. The abnormal species correspond to partial truncation of exon 3 presumably due to the nucleotide change in exon 3, which constitutes a new consensus acceptor splice site within this exon. We postulate that the observed blank is essentially the consequence of qualitative change in a critical region of this novel antigen as abnormal mRNA species are relatively less abundant than normal species. Because the residue 96 of the HLA class I heavy chain is directly involved in interaction with α 2 m, another interesting possibility is that an aminoacid change in this position would perturb such interaction and consequently could affect the serological specificity of B∗3916, or its expression or both.

The CD1 molecules exhibit characteristics of the MHC class I and class II molecules. They are expressed on cortical thymocytes and, similarly to MHC class II molecules, on antigen-presenting cells. In the present study, we investigated the role of the CD1 molecules in the T-cell response to bacterial superantigens. Indeed, we have observed that CD1 molecules could be detected on the CD14-positive population of some healthy donors (14% of donors tested). The CD1 expression on monocytes is correlated with an activation state of the donors as demonstrated by the increased expression of the CD25, CD38, CD45R0, and MHC class II molecules on their lymphocytes. On these donors, CD1a mAbs induced a clear inhibition (65%) of lymphocyte proliferation induced by either staphylococcal enterotoxin A or toxic shock syndrome toxin-1, whereas this proliferation was constantly unaffected by the addition of mAbs directed against CD1b or CD1c. Moreover, an intracellular calcium flux was induced in monocytes following CD1a engagement, and this calcium flux was partially inhibited by preincubation of these cells with the superantigen. These results attribute to the CD1a molecule expressed by monocytes a role in the transduction of signal(s) involved in superantigen-induced activation.

In this study, we used the affected sibling-pairs approach to investigate the linkage of HLA (human leukocyte antigen)-DRB* with phenotypes related to allergy to Parietaria, the most common pollinosis in Mediterranean countries. The study population consisted of 51 nuclear families (235 subjects). Linkage was detected with Parietaria skin test positivity (p(0.01), presence of IgG and IgE antibodies specific for the major allergen Par o 1 (p0.020 and p0.025, respectively), and absence of Par o 1-specific IgE (p0.020). High levels of Par o 1-specific IgG were associated with DRB1*1101 and/or DRB1*1104 (p0.0001 and p0.0119, respectively) in parents and probands. High levels of Par o 1-specific IgE were associated with DRB*1104 in parents (p0.017) and with DRB1*1101 in probands (p0.0146). When siblings were categorized according to high/low total IgE levels (or =125 IU/ml and125 IU/ml, respectively), high IgE antibody response was associated with DRB1*1104 in siblings with low total IgE (p0.034) and with DRB1*1101 in siblings with high total IgE (p0.05). These results demonstrate that HLA-DRB1*, or genes in linkage disequilibrium, contributes to susceptibility to Parietaria allergy and that total IgE levels can discriminate population subsets where different alleles (at the HLA region or at loci in linkage disequilibrium) contribute to control allergen-specific IgE synthesis.

Cord blood is increasingly used in transplantation as it is a readily available source of progenitor cells and is reputed to generate less severe graft-versus-host disease (GVHD) than adult bone marrow. We have compared apoptosis of cord blood lymphocytes (CB) and adult lymphocytes (PBMC) after stimulation via HLA class I, HLA class II or CD3 in order to reproduce in vitro some of the stimuli occurring after allotransplantation. CB spontaneously apoptose more than PBMC ex vivo, stimulation via HLA class I dramatically increased CB apoptosis without altering viability of PBMC. Expression of Fas was markedly lower on CB than on PBMC and this difference was maintained even after activation. Fas ligand was expressed in CB and in PBMC. CB were activated via either HLA class I or class II molecules although proliferation was not observed. Only phorbol ester pre-activation allowed Fas to subsequently induce a death signal. Proliferation of PBMC via CD3 led to enhanced Fas signals. CB therefore differ from PBMC with regard to both spontaneous and activation induced apoptosis and either allo- or CD3 mediated stimulation. Finally, the apoptosis of CB via HLA-class I could have an important role in the moderation of graft-versus-host disease.

In this report we show that two widely-used monoclonal antibodies, TAL-1B5 and DA6.147, which react with the HLA-DR alpha chain on immunoblots, recognize the C-terminal intracellular tail of this HLA-DR subunit. We demonstrate that both MoAbs react with a synthetic peptide representing the intracellular C-terminal tail of the DR alpha chain and that mutant DR molecules lacking this part of the alpha chain lose reactivity with TAL-1B5 and DA6.147, both in Western blot analysis and in intracellular FACS staining.

Research on stem cell therapies for regenerative medicine is progressing rapidly. Although the use of autologous stem cells is a tempting choice, there are several instances in which they are either defective or not available in due time. Allogenic stem cells derived from healthy donors presents a promising alternative. Whether autologous or allogenic, recent advances have proven that stem cells are not as immune privileged as they were thought. Therefore understanding the interactions of these cells with the recipient immune system is paramount to their clinical application. Transplantation of stem cells induces humoral as well as cellular immune response. This review focuses on the humoral response elicited by stem cells upon their administration and consequences on the survival and maintenance of the graft. Current transplantation identifies pre- and post-transplantation anti-HLA antibodies as immune rejection and cell signaling effectors. These two mechanisms are likely to operate similarly in the context of SC therapeutics. Ultimately this knowledge will help to propose novel strategies to mitigate the allogenic barriers. Immunogenetics selection of the donor cell and immunomonitoring are key factors to allow the implementation of regenerative stem cell in the clinics.

Interleukin 1 (IL-1) exerts biological properties on various immune and nonimmune cell types and tissues and thus may play an important role during chronic inflammatory processes. Here we have examined the IL-1 biosynthesis in adherent synovial lining cell (ASLC) cultures obtained from patients with rheumatoid arthritis (RA). We report that ASLCs in culture showed heterogeneous endogenous levels of IL-1 alpha and beta expression. Recombinant interleukin 1 (rIL-1) alpha or beta induced increases of IL-1 alpha and beta mRNA and proteins levels in ASLCs. Although IL-1 synthesis is enhanced by rIL-1 treatment, no soluble IL-1 alpha or beta could be detected by specific enzyme-linked immunosorbent assays. A pretreatment with recombinant IFN gamma (rIFN gamma) down-regulated the effect of rIL-1 on IL-1 synthesis in ASLCs. Actinomycin D suppressed the endogeneous and rIL-1-induced IL-1 mRNA expression Indomethacin, in the presence of rIL-1 alpha or beta, up-regulates the level of expression of IL-1 beta in ASLCs pretreated with rIFN gamma, but has the opposite effect in non-pretreated cells. The increase of IL-1 gene expression by rIL-1 in human ASLCs from RA patients may contribute as an amplification of the disease progress. These studies may also explain the beneficial effects of IFN gamma in experimental models of IL-1-induced bone and cartilage degradation and in patients with diseases involving IL-1.

Major histocompatibility complex (MHC) class I chain-related A (MICA) molecules mediate natural killer (NK) cell activation and T lymphocyte co-stimulation. A polymorphic methionine (met) to valine (val) variation at amino acid position 129 of the alpha2 heavy chain domain is in linkage disequilibrium with other allelic changes and seems to categorize MICA alleles into strong and weak binder of NKG2D receptor and thereby to influence effector cell function. We investigated here whether MICA-129 dimorphism is associated with susceptibility to/or resistance against developing nasopharyngeal cancer (NPC). DNA from 130 NPC patients and 180 healthy individuals from Tunisia were genotyped for MICA-129 variation. We found a higher frequency of MICA-129 val/val genotype in patients than in controls (corrected p value = 0.02) that could suggest a tumor escape mechanism because of failure to activate NK cells by MICA-129 val allele or absence of NK cell activation because of absence of MICA-129 met allele in individuals otherwise predisposed to viral/environmental factors.

Aim Today, using human cardiac-derived stem/progenitor cells (hCPC) to repair injured inflamed myocardium is beyond cell stemness-related biologics, and involves paracrine effect as important therapeutic element in addition to regenerative one. Autologous hCPC were proving not to be optimum. This enhanced the prospects of using allogeneic cells although allogenicity is a striking system that can produce detrimental and beneficial effects. We aimed at defining whether allogeneic setting of hCPC within the context of cardiac repair is detrimental or beneficial. Methods Using an experimental model of allogeneic stimulation we investigated hCPC crosstalk with adaptive and innate immune effector cells and their reactivity with allo-HLA antibodies. Results Whether under inflammatory conditions or not, interactions of hCPC with major composites of the adaptive and innate immune system, T and natural killer cells, respectively, in allogeneic setting trigger and expand regulatory anti-inflammatory rather than inflammatory/deleterious immune response. The HLA molecules but also immune-regulatory molecule PD-L1 orchestrate this “tolerogenic/immunomodulatory” behavior. In regard of humoral antibody-mediated immunity, hCPC can variably react with allo-HLA antibodies present in serum depending on their HLA expression level. hCPC reactivity with anti-HLA resulted in low complement-dependent cytotoxicity and negligible antibody-dependent cell-mediated cytotoxicity thus, minimal deleterious effect. In contrast, we found that engagement of HLA class I or II molecules on hCPC by respective specific antibodies induces rapid phosphorylation of a major pathway of cells proliferation and survival, MAPK/Erk. Although warrant further investigation, this suggests that allo-HLA antibodies could enhance engraftment, proliferation, and survival of hCPC and potentially contribute to myocardium repair. Conclusions Collectively, our data indicate that allogenicity of hCPC within cardiac repair context is beneficial rather than detrimental. It promotes the immune response toward regeneration/reparation and suggests allogeneic hCPC crosstalk with composites of immune system as an eventual “paracrine” therapeutic element.

Major histocompatibility complex (MHC) class I chain-related A (MICA) molecules deliver activating signals through the NKG2D receptor expressed on the surface of natural killer (NK), CD8alphabeta and gammadelta T cells, and the MICA gene is polymorphic. The recently described MICA amino acid substitution at position 129 (MICA-129) seems to affect its binding to NKG2D. We investigated whether this dimorphism (MICA-129met [methionine] and MICA-129val [valine]) is associated with susceptibility to ankylosing spondylitis (AS) in a cohort of Algerian patients stratified according to their HLAB27 status and the age of onset of the disease. DNA from 129 patients and 76 healthy individuals were analyzed to determine the HLA-B generic type as well as MICA-129 polymorphism. Statistical analysis revealed: (1) a weaker association between AS and HLA-B27 in Algerians than in that reported for European patients (63% versus 80-90%), suggesting a possible influence of other genetic/environmental determinants in the studied population and (2) an association between MICA-129 met/met genotype and juvenile AS (p = 0.02) independent of HLA-B27 status. These data suggest a potential role for a functionally relevant MICA gene polymorphism in autoimmune/inflammatory disease susceptibility.

Despite systematic antibiotic therapy, severe infections (septicemia, meningitis, or osteomyelitis) are a major cause of mortality and morbidity in children with sickle cell disease (SCD). In this study, we explored the possibility that polymorphism at the human leukocyte antigen (HLA) locus might constitute an immunogenetic modifying factor to the intrinsic susceptibility to infection in patients with SCD. A cohort of 80 SCD patients living in Paris, 43 with at least one major infectious complication and 37 without infections, were typed for HLA class II loci by polymerase chain reaction-sequence-specific primers (PCR-SSP). We found that significantly more patients without infections carry the HLA class II DRB1∗15 specificity than did patients with infections (21.6% in the first group, versus 4.7% in the second group; χ 2 = 10.47, p c = 0.01), supporting a protective effect of this allele. Conversely, significantly more patients were found to carry the DQB1∗03 specificity within the group of severe infections, supporting a negative effect (34.9% versus 12.2%, χ 2 = 9.41, p c = 0.01). These findings suggest a direct involvement of HLA polymorphism in the development of major infections in SCD. Together with previous data on polymorphism of the Fc receptor and of the mannose-binding lectin, they provide evidence for a polygenic immunomodulation of the constitutively increased infectious risk in SCD.

Primary Sjogren syndrome (pSS) is an autoimmune disease characterized by progressive destruction of the exocrine glands leading to mucosal and conjonctival dryness. It is marked by lymphocytic infiltration of the glands and the accumulation of several types of autoantibodies such as rheumatoid factor (RF), antinuclear, anti-SS-A (anti-Ro) and anti-SS-B (anti-LA) autoantibodies. The susceptibility to pSS and/or the presence of SS-A/SS-B autoantibodies in pSS patients is associated with DRB1∗03-DQB1∗02 and DRB1∗02-DQB1∗06 haplotypes, whereas no associations have been described with any HLA class I allele. To define the impact of HLA class I alleles in predisposition to pSS, 46 patients responding to the European criteria and 222 healthy unrelated Caucasians were analyzed for their HLA class I and class II haplotypes. Our results confirm the association of the DRB1∗03-DQB1∗02 haplotype with SS-A/SS-B autoantibodies positive pSS and demonstrate a significant association of the HLA-A24 with the disease. Moreover, HLA-A24 is more often associated with DRB1∗11-DQB1∗0301 and/or DRB1∗0301-DQB1∗02 in pSS patients than in the controls. The novel association of HLA class I alleles with susceptibility to pSS provides new insights to the genetic predisposition to this disease and subsequently to its physiopathology.

In addition to lymphocyte phenotyping and functional assays, new tools are now available to monitor specific aspects of the immune response in the follow-up of hematopoietic stem cell transplantation: reconstitution of T-cell diversity (spectratyping or immunoscope), ex vivo thymic function by measuring "T-cell receptor rearrangement excision DNA circles" and antigen-specific T-cell responses (HLA tetramers). Combining these methods should contribute to improve our current knowledge of how the immune system is reconstituted in different settings of hematopoietic stem cell transplantation and how we could improve this recovery process.

Bacterial superantigens (Sag) are potent activators of T cells. This T-cell activation has been described as an MHC class II dependent phenomenon. We have observed that human thymocytes depleted of MHC class II positive cells are still able to proliferate in response to the staphylococcal enterotoxin A (SEA). This proliferation was clearly inhibited by the addition of monoclonal antibodies directed against the CD1a molecule. In contrast, monoclonal antibodies directed against the CD1b and CD1c molecules have no effect on the Sag-induced activation of the CD2 (+) MHC class II (−) thymocytes. We next examined the ability of the CD1a molecule to transmit transmembrane signals. Results obtained indicate that CD1a ligation on these thymocytes induced tyrosine phosphorylation of the p56 lck tyrosine kinase. Signal transduction via CD1a is further confirmed by the observation of a significant intracellular calcium flux (Ca i ++ ) in thymocytes following CD1a engagement. These data demonstrate that CD1a ligation induces a signal transduction pathway which has a potential role in the bacterial superantigen-induced activation of human CD2 (+) MHC class II (−) thymocytes.

Hematopoietic progenitors express HLA-DR molecules. However the significance of HLA-class II molecules on CD34+ cells remains unknown. The primary role of HLA-class-II molecules is antigen presentation although a second role, that of signal transduction, has been established in B cells. The role of HLA-DR in hematopoiesis was examined by determining the ability of CD34+ progenitor cells to differentiate to “Colony Forming Unit Granulocyte-Macrophage” (CFU-GM) and “Burst Forming Unit Erythrocyte” (BFU-E) in the presence of anti-HLA-DR monoclonal antibody. We observed a reduction in the number of CFU-GM which was due in part to down regulation of granulocyte rather than monocyte differentiation. These observations suggest that HLA-DR signals can regulate myelopoiesis. We point out especially the role of the HLA-DR molecule in the switch of CFU-GM between granulocyte or monocyte lineages. Although HLA-DR mediated apoptosis has been described in mature B lymphocytes apoptosis of CD34+ cells was excluded as a mechanism.

Cord blood is increasingly used in transplantation as it is a readily available source of progenitor cells and is reputed to generate less severe graft-versus-host disease (GVHD) than adult bone marrow. We have compared apoptosis of cord blood lymphocytes (CB) and adult lymphocytes (PBMC) after stimulation via HLA class I, HLA class II or CD3 in order to reproduce in vitro some of the stimuli occurring after allotransplantation. CB spontaneously apoptose more than PBMC ex vivo, stimulation via HLA class I dramatically increased CB apoptosis without altering viability of PBMC. Expression of Fas was markedly lower on CB than on PBMC and this difference was maintained even after activation. Fas ligand was expressed in CB and in PBMC. CB were activated via either HLA class I or class II molecules although proliferation was not observed. Only phorbol ester pre-activation allowed Fas to subsequently induce a death signal. Proliferation of PBMC via CD3 led to enhanced Fas signals. CB therefore differ from PBMC with regard to both spontaneous and activation induced apoptosis and either allo- or CD3 mediated stimulation. Finally, the apoptosis of CB via HLA-class I could have an important role in the moderation of graft-versus-host disease.

Although the production of selected cytokines by B cells is important for their regulation, little is known about MHC class II-induced cytokine expression in these cells. We designed the present studies to investigate MHC class II-mediated TNF-beta gene expression in 19 EBV-transformed homozygote B cell lines at similar stage of differentiation but presenting different MHC class II haplotypes. Our results demonstrate that in contrast to PMA, engagement of MHC class II with staphylococcal enterotoxin A (SEA), a natural ligand, or with anti-HLA-DR mAb L243, stimulates TNF-beta gene expression in some but not all B cell lines. The differential stimulation of TNF-beta gene expression via MHC class II was not due to the cells MHC class II expression level, nor to their capacity to bind the ligands as evidenced by SEA binding affinity studies. Together these results demonstrate that ligation of MHC class II molecules can stimulate TNF-beta gene expression in a B cell line-dependent manner. The differential cytokine gene expression might be due to an influence of MHC class II haplotype either by a linkage disequilibrium with TNF-beta gene or by a differential association with effector or cell surface molecules.