Twenty-seven cord blood samples from healthy newborns were processed according to a "whole blood" flow cytometric analysis. The CD3-positive T cells were a variable subpopulation representing 44.8 +/- 13.3% of lymphocytes. The majority of the CD3+ cells are CD38+. Newborn T cells have lower levels of both IL-2 receptors and HLA-DR than do adult T cells. The CD4-positive T cells represented 31.0 +/- 10.8% of lymphocytes with a great prevalence of the CD4+/CD45RA+ population. The CD3+/CD8+/CD11b+ cells are increased to 23.4 +/- 7.1% of lymphocytes. The CD57 antigen is not expressed. The NK population, CD16+/CD56+, is increased to 25 +/- 11% of lymphocytes. Of CD19+ cord blood B lymphocytes 68% coexpressed CD5. Thus "suppressive" and "naive" cells are prominently represented in cord blood.
Fetal mononuclear cells are increasingly used in transplantation of hematopoietic cells due to a reportedly lower incidence of graft-versus- host disease. Previous studies of immune responses of fetal lymphocytes have indicated a general hyporesponsiveness in response to polyclonal stimulation. Fetal B lymphocytes display many features typical of the resting state such as a low level of HLA class II expression, but a large proportion of cells also carry the activation-associated CD23 antigen. We show here that despite a low cell surface level of all three HLA class II isotypes on fetal B cells, their allogeneic capacity, measured as the ability to elicit a mixed lymphocyte reaction, is similar to that of adult B cells. Allogeneic stimulation is believed to be peptide-dependent. Surprisingly, the majority of the HLA class II molecules on cord blood B cells appeared to be devoid of stably bound peptide as detected by the binding of a recombinant and soluble invariant chain, as well as by the absence of sodium dodecyl sulfate (SDS) stable alpha beta heterodimers in whole cell lysates. Immunoblot experiments showed that HLA class II molecules of fetal B cells were predominantly present in high molecular weight aggregates in stark contrast to B cells of adult origin. However, a sensitive cell surface labeling technique followed by immunoprecipitation enabled us to detect an SDS-stable 120-kD molecule on fetal B cells. We propose that the 120-kD molecules could correspond to HLA class II doubledimers or superdimers. We hypothesize that the 120-kD HLA class II molecule functions as the antigen-presenting molecule in the mixed lymphocyte reaction of fetal B cells, as it is the major species detected on the surface. Secondly, we suggest that a high level of empty HLA class II molecules may be indicative of a particular stage in B-cell ontogeny.
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