J.J. van Rood

Simultaneous detection of two cell populations by two-colour fluorescence and application to the recognition of B-cell determinants.

CELL separation techniques, the possibility of recognising different markers by immunofluorescence and functional tests, have subdivided cells once thought to be a homogeneous group into different subclasses. This development has been most prominent for the so-called mononuclear cells. A difficulty inherent in the classification of such subclasses (of, for example, mononuclear cells) is that it is often almost impossible to determine precisely what percentage of the cells belongs to a certain subclass, and whether the sum of the subclasses is equal to, lower than or even higher than 100% of the total of the cells analysed. A typical recent example was the identification of the “null” or K cell1,2. Such analysis is greatly facilitated if we analyse a cell suspension stained with two or more different fluorescent dyes visible at the same time. This is feasible.

Results of allogeneic bone marrow transplants for leukemia using donors other than HLA-identical siblings.

PURPOSE To compare outcomes of bone marrow transplants for leukemia from HLA-identical siblings, haploidentical HLA-mismatched relatives, and HLA-matched and mismatched unrelated donors. PATIENTS A total of 2,055 recipients of allogeneic bone marrow transplants for chronic myelogenous leukemia (CML), acute myelogenous leukemia (AML), and acute lymphoblastic leukemia (ALL) were entered onto the study. Transplants were performed between 1985 and 1991 and reported to the International Bone Marrow Transplant Registry (IBMTR). Donors were HLA-identical siblings (n = 1,224); haploidentical relatives mismatched for one (n = 238) or two (n = 102) HLA-A, -B, or -DR antigens; or unrelated persons who were HLA-matched (n = 383) or mismatched for one HLA-A, -B, or -DR antigen (n = 108). HLA typing was performed using serologic techniques. RESULTS Transplant-related mortality was significantly higher after alternative donor transplants than after HLA-identical sibling transplants. Among patients with early leukemia (CML in chronic phase or acute leukemia in first remission), 3-year transplant-related mortality (+/-SE) was 21% +/- 2% after HLA-identical sibling transplants and greater than 50% after all types of alternative donor transplants studied. Among patients with early leukemia, relative risks of treatment failure (inverse of leukemia-free survival), using HLA-identical sibling transplants as the reference group, were 2.43 (P < .0001) with 1-HLA-antigen-mismatched related donors, 3.79 (P < .0001) with 2-HLA-antigen-mismatched related donors, 2.11 (P < .0001) with HLA-matched unrelated donors, and 3.33 (P < .0001) with 1-HLA-antigen-mismatched unrelated donors. For patients with more advanced leukemia, differences in treatment failure were less striking: 1-HLA-antigen-mismatched relatives, 1.22 (P = not significant [NS]); 2-HLA-antigen-mismatched relatives, 1.81 (P < .0001); HLA-matched unrelated donors, 1.39 (P = .002); and 1-HLA-antigen-mismatched unrelated donors, 1.63 (P = .002). CONCLUSION Although transplants from alternative donors are effective in some patients with leukemia, treatment failure is higher than after HLA-identical sibling transplants. Outcome depends on leukemia state, donor-recipient relationship, and degree of HLA matching. In early leukemia, alternative donor transplants have a more than twofold increased risk of treatment failure compared with HLA-identical sibling transplants. This difference is less in advanced leukemia.

TRANSPLANTATION OF BONE-MARROW CELLS AND FETAL THYMUS IN AN INFANT WITH LYMPHOPENIC IMMUNOLOGICAL DEFICIENCY

Abstract A 5-month-old male infant with lymphopenic immunological deficiency (Swiss-type agammaglobulinaemia) was treated by transplantation of a fetal thymus, and of bone-marrow-derived cells from his 7-year-old sister. The bone-marrow donor was selected for histocompatibility by leucocyte typing at the HLA locus and by mixed lymphocyte culture. Nucleated bone-marrow cells were centrifuged on an albumin gradient and a small number (5 × 10 6 /kg. body-weight) from the fraction assumed to contain concentrated haemopoietic stem cells, were transplanted. 3 months after the transplantation the boy resumed normal development, and the cellular and humoral immunity were restored. There was an increase in the three major classes of immunoglobulins to a level that exceeded normal adult values. In addition a homogenous IgG-L and an IgA-K band were detected in the serum. A temporary mild exanthema was taken to be a sign of graft-versus-host reaction. Nursing in a laminar-flow cabinet appeared to be relatively simple and highly effective for protection against contamination with microorganisms from the environment.

HLA-DR phenotypes in Spanish coeliac children: their contribution to the understanding of the genetics of the disease.

The DR-locus controlled B-cell antigens were studied in 163 unrelated Spanish coeliac children and 68 families of this group, nine of them with more than one coeliac patient, to obtain more information about the association between these antigens and coeliac disease. The results show that the most common coeliac phenotypes are DR3/DR7, DR7/DR5, DR3/other DR, and DR3/DR3. The family study confirmed the segregation of the disease with the above mentioned phenotypes. In eight of the nine multiple case families, all coeliac children shared both HLA-DR antigens. These findings make it unlikely that a single dominant gene linked to HLA-DR controls the susceptibility to coeliac disease. The phenotypes in the patients were not distributed according to the Hardy-Weinberg equilibrium. Thus, a model based on one recessive susceptibility gene linked to HLA-DR is not probable either. The complexity of the genetics of coeliac disease and some of the features shared with the HLA-DR pattern in juvenile insulin-dependent diabetes are discussed.

HLA-UNKED GENETIC CONTROL OF HOST RESPONSE TO MYCOBACTERIUM LEPRÆ

Abstract Non-random parental HLA-haplotype segregation is demonstrated in siblings with leprosy. A new method is described for the statistical analysis of non-random segregation among sibships of different sizes. Sibs with the same type of leprosy show a significant excess of identical HLA haplotypes. This is also true for families in which only tuberculoid leprosy is found, which is by far the commonest type in the population studied. However, sibs affected with different types of leprosy share a haplotype less often than expected. This indicates that both susceptibility to and type of leprosy are controlled by at least two HLA-linked genes. Our findings suggest that the equivocal results of previous population studies are due to differences of linkage disequilibrium between HLA-linked genes controlling the host response to Mycobacterium leprœ and alleles of HLA A and B loci in various populations.

HLA-DW3 ASSOCIATED WITH CŒLIAC DISEASE

28 patients with coeliac disease (C.D.) were typed for the HLA-A, -B, and -D loci by several techniques. It was found that C.D. is primarily associated with the DW3 determinant and only secondarily with HLA-B8. The previously described association with HLA-B8 is explained by linkage disequilibrium between HLA-B8 and DW3.

Rejection of bone-marrow graft by recipient-derived cytotoxic T lymphocytes against minor histocompatibility antigens.

A female patient showed rejection of a T-lymphocyte-depleted bone-marrow graft from her phenotypically HLA-identical father. Before bone-marrow transplantation, there was strong recipient anti-donor cellular cytotoxic reactivity directed against several minor histocompatibility (mH) antigens, including the male-specific H-Y antigen. After conditioning treatment, no recipient anti-donor cytotoxic activity could be detected, and good graft function was shown a month after transplantation. Thereafter, however, graft function deteriorated rapidly, while recipient-derived anti-donor cellular cytotoxic reactivity, against similar mH antigens, reappeared. The recipient-derived cytotoxic T lymphocytes could completely inhibit growth of donor haemopoietic progenitor cells both before and after bone-marrow transplantation. Thus, cytotoxic T lymphocytes can survive very intensive conditioning regimens, and residual recipient cytotoxic T lymphocytes directed against mH antigens expressed on donor haemopoietic progenitor cells may cause graft rejection after HLA-identical T-lymphocyte-depleted bone-marrow transplantation.

Gluten Sensitive Enteropathy in Spain: Genetic and Environmental Factors

It has been postulated by various investigators that at least two unlinked genes are involved in the predisposition to gluten sensitive enteropathy (GSE). One of these genes is believed to be linked to the major histocompatibility locus (MHC). We call this gene GSE1 and the other, which is not linked to MHC, GSE2. The study of HLA antigens in different populations may help to elucidate the genetics of GSE and to identify the GSE1 gene. We have studied 26 unrelated proven GSE children and 54 controls living in a defined region in Spain, and also performed a family study in 51 relatives of 11 of these children with GSE. These relatives also underwent a physical examination and a routine haematological screening, and in 30 of them a small-intestinal biopsy specimen was investigated. All patients, relatives, and controls were typed for HLA-A, B, C and DR antigens and for glyoxalase-1 (GLO1). Both the population and the family studies have shown that GSE is associated with two alleles at the HLA-DR locus, namely HLA-DRw3 and HLA-DRw7. No association with a particular GLO1 allele was found. Three monozygotic twin pairs, one concordant and two discordant for the disease, are described. In addition, all monozygotic twin pairs in the literature are reviewed and several obtained through personal communication are added.

Genetic control of survival in epidemics.

Descendants of Dutch colonists, who emigrated to Surinam in the last century and survived epidemics of typhoid and yellow fever with a total mortality of about 60%, were tested for twenty‐six polymorphisms. The gene frequencies were compared with those of a large Dutch control sample. An analysis of drift indicated that the variations in gene frequencies observed for C3, Gm, HLA‐B, and GLO were unlikely to be due to drift. Therefore these data might indicate selection through genetic control of survival in these epidemics.

MATCHING FOR HLA ANTIGENS OF A, B, AND DR LOCI IN RENAL TRANSPLANTATION BY EUROTRANSPLANT

79 patients and their respective cadaveric kidney donors were typed for HLA-A, HLA-B, and HLA-DR antigens using frozen stored spleen lymphocytes and fresh peripheral-blood lymphocytes. The kidney-graft survival-rate at 3 to 18 months was highest when donor and recipient shared one or two DR antigens and three or four A and B antigens. The graft-survival rate was significantly higher (87 +/- 6%) at 18 months in these patients than in less well matched patients (48 +/- 9%).