There are two C4 genetic loci and a null allele in the chimpanzee.

Genetic polymorphism in C4 in the chimpanzee was studied by agarose gel electrophoresis of desialated plasma and development of patterns by immunofixation with antiserum to human C4 and by a C4-sensitive hemolytic overlay. In general, immunofixation patterns showed multiple partially overlapping bands of which only the most cathodal had strong hemolytic activity. In analogy to human C4, the latter were designated C4B, whereas those detected by immunofixation which had little hemolytic activity were designated C4A. Chimp C4A and C4B reacted with human and mouse (monoclonal) anti-C4B and human anti-Ch1 but neither reacted with monoclonal anti-C4A or human anti-Ch2, Ch3, Rg1, or Rg2. On sodium dodecyl sulfate polyacrylamide gel electrophoresis, the alpha chain of C4B showed a slightly lower apparent relative mass than that of C4A at around Mr 93,000. There were three C4A variants and two C4B variants inherited in families as autosomal codominant traits, as C4A-C4B cosegregating pairs with no detectable crossing-over. These pairs were inherited with chimpanzee leukocyte antigen types C2 and BF variants without detectable crossing-over. Half-null C4 haplotypes with C4B QO were observed in family studies. Nine BF, C2, C4A, C4B allelic haplotypic combinations (complotypes) were identified among presumably unrelated chimpanzees.

Differentiation antigens on rhesus monkey lymphocytes. II. Characterization of RhT3, a CD3-like antigen on T cells.

A monoclonal antibody FN18 is described, which is specific for mature rhesus monkey T lymphocytes. It defines a cell surface antigen, composed of two polypeptide chains with a molecular mass of 22 and 27 kDa. In view of these and other similarities with the human T3 or CD3 antigen, it was designated as RhT3. Expression, distribution and certain functions of RhT3 were compared with those of its human counterpart. Analogous to anti-CD3 antibodies in man, FN18 is able to modulate its target antigen, has mitogenic properties and is able to block the pokeweed mitogen and concanavalin A-driven cell proliferation, but not that caused by phytohemagglutinin. In spite of such minor dissimilarities the available data strongly suggest that the RhT3 antigen is the rhesus monkey's homologue of the human CD3 antigen.

Differentiation antigens on rhesus monkey lymphocytes. I. Identification of T cells bearing CD3 and CD8, and of a subset of CD8-bearing cells.

Rhesus monkeys provide an excellent preclinical model to test the effect of monoclonal antibodies (mAb) in vitro and in vivo. So far, mostly mAb have been used which were originally raised against human cell surface antigens but cross-reacted reasonably well with homologous antigens on rhesus monkey cells. However, to optimize the model, it was necessary to produce mAb which react specifically with subsets of rhesus monkey lymphocytes. In this report, three mouse anti-rhesus monkey mAb are described, specific for different subsets of rhesus monkey T lymphocytes. None of the reagents cross-reacts with human lymphocytes. Characterization of these mAb was based upon indirect immunofluorescence, using a simultaneous staining technique, and immunoprecipitation of the specific target antigens. One antibody (GM9) reacts with the same subset as is recognized by mAb specific for human CD8+ cells. The second mAb (GM13) is specific for a subset of CD8+ cells. A third mAb (FN18) was of particular interest: it identifies a cell surface complex, RhT3, expressed on mature T lymphocytes, of which the polypeptide chains have a molecular mass of 22 and 27 kDa. The data strongly suggest that RhT3 is a CD3-like determinant, so far unidentified in the rhesus monkey.

Recommendations for the housing of macaque monkeys.

A multidisciplinary working group was formed to make recommendations for housing of macaques under laboratory conditions in the Netherlands. The group concluded that long-term individual caging leads to persistent abnormal behaviour. Therefore, individual housing is regarded as acceptable only for special reasons which counter-balance the adverse effects of isolation. Guidelines are given for developing more satisfactory social housing systems. Cages used in individual as well as social housing should meet certain spatial and other requirements to ensure a certain amount of diversion, freedom of movement and safety. Since the recommendations represent the opinion of experts in certain aspects of animal husbandry, the report can be used as a legal reference under the Animal Experiments Act.

Immune response against vaccinia virus in rhesus monkeys: no evidence for primary MHC-restricted cytolytic T cells.

Rhesus monkeys were tested in vitro for their cellular immune response after infection with vaccinia virus, employing lymphocyte preparations from various lymphoid tissues. Although virus-infected target cells were lysed by lymphoid cells from immunized, but not from uninfected, rhesus monkeys, we could neither find evidence for MHC-restricted T cells nor for antibody-dependent cellular cytotoxicity. Kinetics of target cell lysis, the killing patterns of immune lymphocytes measured on syngeneic, allogeneic, and xenogeneic target cells, and the influence of protein A on cytotoxic activity in vitro suggest induction predominantly of natural killer cells in vivo which exhibit lytic activity on virus-infected target cells in vitro.

Platelet and granulocyte specific allo-antigens in chimpanzees.

Chimpanzees were allo-immunized with blood from donors, ChLA-A and -B identical to the recipient. The sera of some of these animals contained antibodies that reacted only with the platelets or granulocytes of the immunizing donor and of some related and unrelated chimpanzees. Both for the platelets and the granulocytes a di-allelic system of allo-antigens is described, provisionally called ChPL-1 and ChGR-1.

Effects of in vivo administration of monoclonal antibodies specific for human T cell subpopulations on the immune system in a rhesus monkey model.

Monoclonal antibodies specific for human T cell subsets have been tested for their immunosuppressive effect in a rhesus monkey skin graft model. Rhesus monkeys were injected i.v. daily with antibodies specific for helper T cells (OKT4 and 4A), for cytotoxic/suppressor T cells (OKT8A), or all peripheral T cells (OKT11A), and they received an allogeneic skin graft one or two days after the initial antibody treatment. The OKT4, 4A, and 11A antibodies prolonged skin graft survival, but OKT8A did not. All animals were carefully monitored regarding levels of T cell subsets and antibody formation to the injected monoclonal antibody. The relevant T cell subset was not eliminated from the circulation when OKT4 and OKT4A antibodies were given separately. The OKT4+ cells remained in the circulation coated with antibody. OKT4+ cells could no longer be demonstrated when both OKT4 and 4A were given simultaneously. However, this difference in effect on the OKT4+ cells did not influence skin graft survival time. All animals receiving monoclonal antibody treatment developed antimouse-Ig antibodies after 10 to 13 days of treatment, which presumably counteracted the effect of the antibodies. From these data it appears that the rhesus monkey is a useful animal model in which to investigate the potential of monoclonal antibodies against human lymphocyte subpopulations to modify and regulate the immune response in an orderly fashion.

The major histocompatibility complex of rhesus monkeys, RhLA: XV. Chemical characterizations of DR locus antigens and antigen 48.

Immunoprecipitation studies of the rhesus monkey major histocompatibility system have shown that the RhLA-DR locus codes for class II antigens with molecular features that are homologous to the class II antigens coded for by the human HLA-DR locus. The product of another alloantigenic RhLA-linked locus of the rhesus monkey, called '48', is provisionally characterized as a class I system.

Kidney transplantation in rhesus monkeys. Matching for D/DR antigens, pretransplant blood transfusions, and immunological monitoring before transplantation.

The effect of matching for D/DR antigens and of three pretransplant blood transfusions on kidney allograft survival was investigated in unrelated rhesus monkeys treated with standard immunosuppression. A control group consisting of host-donor combinations mismatched for one or two DR antigens (mixed lymphocyte culture (MLC) positive) and not receiving transfusions showed a MST of 13 +/- 1.2 days with a range from 9 to 22 days. The administration of pretransplant blood transfusions led to a MST of 28 +/- 5.4 days with 5 of 12 animals showing survival times of more than 22 days (i.e., a bimodal distribution of survival times). Recipients matched with their donors for two DR antigens and given transfusions showed an even better MST of 39 +/- 4.0 days. Under these conditions, MLC-negative combinations fared slightly better than MLC-positive ones: only 1 of 10 animals showed a survival time of less than 22 days and kidney function in the first weeks after transplantation was significantly better. When mixed lymphocyte reactions (MLC reactivity) between host and donor before and after the transfusions were compared, it was possible to predict to some extent the eventual fate of an allograft: increased mixed lymphocyte reaction predicted relatively short survival times, decreased mixed lymphocyte reaction relatively long ones (P less than 0.01).

The technique of kidney transplantation in rhesus monkeys.

The technique of kidney transplantation in rhesus monkeys is described in detail. The rate of successful transplantations can be increased to 90-100% if several technical aspects are considered. These include adequate perfusion of the organ to be preserved, measures to prevent a sharp decrease in body temperature, doing the arterial anastomosis with interrupted sutures and prevention of low blood pressure after revascularization.

Influence of prophylactic treatment with gamma-immunoglobulins on renal function after kidney transplantation in rhesus monkeys.

The effect of prophylactic i.v. administration of high doses of human gamma-immunoglobulin (IgG) on kidney graft survival was investigated in rhesus monkeys treated with azathioprine and prednisolone. In nontransfused recipients not treated with IgG (controls), graft survival ranged from 9 to 22 days; if nontreated animals had been given three pretransplant blood transfusions, graft survival ranged from 9 to 61 days with 42% of the animals showing a prolonged survival time (greater than 22 days). However, in both transfused and nontransfused recipients, the additional pretransplant administration of IgG appeared to have an adverse effect: about 25% of the animals showed accelerated rejection. In addition, serum creatinine levels in IgG-treated recipients were significantly higher on the 3rd day after transplantation than in non-treated monkeys. We concluded that renal transplant patients should be treated with IgG for protection against life-threatening infections only if they have good kidney function.

Typing for RhLA-D in rhesus monkeys: I. Characteristics of ten groups of homozygous typing cells.

Certain characteristics of 38 homozygous typing cell (TC's) of rhesus monkeys were determined. These TC's define ten RhLA-D locus specificities. Eight of them are associated with established RhLA-DR antigens. Two other groups of typing cells, D9 and D10, were previously considered to be associated with "blank" antigens of the DR series; they now appear to be associated with B-cell antigens which are also likely to be controlled by the DR locus. No influence of RhLA-A or B antigens of MLC reactivity was observed. It was shown, however, that products of at least one locus other than D/DR is responsible for MLC stimulation. Whether those MLC antigens are associated with serologically identifiable B-cell antigens which are not controlled by the DR locus, is not yet clear.

Typing for RhLA-D in rhesus monkeys: II. Genetics of the D antigens and their association with DR antigens in a population of unrelated animals.

A population of 94 unrelated rhesus monkeys was typed for MLC antigens using 38 homozygous typing cells which define RhLA-D specificities. A genetic analysis showed that the ten D specificities are alleles of a single locus, the gene frequencies of which are in Hardy-Weinberg equilibrium. The association between the cellularly defined D and the serologically defined DR antigens in the population was usually high, confirming the close relationship between D and DR antigens. Two new associations were found, i.e. between the D-locus antigens 9 and 10 and the serologically defined B-cell antigens 109 and 101, respectively. This observation confirms prior speculations that the latter two antigens may belong to the DR series.

Absence of a correlation between humoral and cellular responses to a vaccinia virus and products of the major histocompatibility complex in rhesus monkeys.

Sixty-two Rhesus monkeys were tested at different times after vaccinia virus infection for virus-specific induction of lymphocyte proliferation in vitro or antibody production in vivo. No association was found between identifiable RhLA-controlled antigens and the strength of the cellular proliferative and/of humoral response.

Pretransplant blood transfusions have an additive positive effect on kidney graft prognosis in D/DR-matched rhesus monkeys.

The effect of blood transfusions on kidney graft survival in D/DR-matched host-donor combinations was investigated in rhesus monkeys treated with azathioprine and prednisolone. If host-donor combinations were mismatched for D/DR and no transfusions were given (controls), graft survival ranged from 9 to 22 days. D/DR matching alone led to survival times of 13 to 66 days with 62% of the animals showing a prolonged survival time ( greater than 22 days). However, when three pretransplant blood transfusions were given in D/DR-matched combinations, the range of graft survival was even better: 19 to 73 days with 90% of the recipients surviving for longer than 22 days. The recipients in this group also had a better kidney function in the first weeks after transplantation than that of the nontransfused D/DR-matched group. Therefore, the beneficial effects of D/DR matching and pretransplant transfusions are additive in the rhesus monkey.

The major histocompatibility complex of primates: evolutionary aspects and comparative histogenetics.

All mammalian species investigated have a chromosomal region designated as the major histocompatibility complex or m.h.c. The biological significance of the m.h.c. goes far beyond controlling the most important histocompatibility or transplantation antigens; the capacity to respond immunologically, the susceptibility to disease (including cancer), the serum level of several complement factors and numerous other biological traits are regulated by genetic systems closely linked within that chromosomal region. While the basic structure of the m.h.c. seems to be rather similar for all mammalian species, the similarities among the m.h.c. of human and non-human primates are particularly impressive. In this communication, m.h.c. gene products of rhesus monkey, chimpanzee and man are compared and reviewed. Evolutionary aspects of the persistence of the m.h.c. region or 'supergene' throughout the animal kingdom are discussed.

The major histocompatibility complex of rhesus monkeys. XIV. A family study of DR and other RhLA-linked cell membrane antigens.

The segregation of B-cell specific (Ia) determinants was studied in a large number of rhesus families. As expected, on the basis of results of a recent population study, the eight serologically defined DR antigens segregated as alleles of a single locus within RhLA. Two or three antigens not controlled by DR remained candidates for a second Ia series closely linked to DR. The 162 identifiable haplotypes provided interesting information regarding positive associations among antigens of two, three or even four sets of RhLA loci. Further, a new series of antigens closely linked to the RhLA-A locus could be postulated. Data obtained from 8 recombinant offspring permitted a better definition of the mapping position of all known loci of RhLA.