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We found that most peripheral CD4 cells co-express a low density of CD8 alpha antigen in African green monkeys (AGM). Further, the cell surface expression of CD4 and the expression of CD4 mRNA underwent a decrease when purified CD4CD8low cells were cultured with mitogen and IL-2. These observations suggest that AGM CD4 cells are subject to loss of CD4 expression after lymphocyte activation. Part of the peripheral CD8 fraction exhibited a significant helper activity which suggested the phenotypic conversion in helper T cells from CD4+ to CD4- in vivo. Simian immunodeficiency virus (SIV) grew well in CD4 panning cells following SIV infection. In contrast, CD4CD8low cells were resistant to SIV infection after their conversion to CD4- cells.

The U5 monoclonal antibody developed by immunizing mice with Japanese monkey lymphocytes could react with lymphocytes of primate species including Old World monkeys, apes, and human. However, the distributions of U5 antigen on major functional subsets of lymphocytes were different in primate species. The U5 antigen was mainly distributed on natural killer (NK) cells in human, but on B cells in Old World monkeys. On the other hand, U5 antigen was detected on both B and NK cells in chimpanzees and gibbons, indicating that the distribution of U5 antigen on lymphocyte might change from B cells to NK cells during primate evolution.

The mitogen (PHA and ConA)-induced lymphocyte blastogenesis was determined with 15 obese cynomolgus monkeys who were classified into spontaneously occurred obese with hyperlipamic and/or diabetes. There was no difference in the blastogenesis induced by PHA and ConA between obese and normal monkeys when their lymphocytes were cultured in medium supplemented with fetal calf serum. The blastogenesis of obese monkey lymphocytes, however, were significantly lower than that of normal monkey in the case of culturing them in the medium supplemented with autoserum. The wide variation was observed in blastogenesis of normal monkey lymphocytes when they were cultured with obese monkey serum, and some of obese monkey sera significantly inhibited blastogenic response of normal monkey lymphocytes. These results indicate that the responsiveness to mitogensin obese monkey lymphocytes might be normal but their serum might inhibit blastogenesis. Then, the relationship was determined between inhibitory effect of serum and contents of several biochemical markers associated with obese. The significant negative correlation was observed between inhibitory effect and total cholesterol as well as low-density cholesterol (VLDLC+LDLC). This result indicates that the level of serum LDLC+VLDLC possibly correlates with the inhibitory effect, and the increase of serum LDLC+VLDLC with obese might depress the lymphocyte function, resulting in the decrease of immune function in obese patients.

Single laser flow cytometry was applied to the karyotype analysis of green monkeys. Clear sex difference in flow karyotype was recognized in this monkey, because Y chromosome could be identified as a single peak in the histogram of male specimens. We could isolate Y chromosome of this species by the use of a cell sorter, and demonstrate by polymerase chain reaction that the sorted-out chromosomes contained the Y chromosome specific nucleotide sequence (SRY). This chromosome sorting technique provides a powerful strategy for constructing the DNA library specific to Y chromosome in this species.

In contrast to the case of peripheral T cells, the surface expression of CD20 antigen and the expression of CD20 mRNA in monkey lymph node (LN) T cells underwent a noticeable increase when they were cultured with mitogen and interleukin-2 (IL-2). To confirm in vivo regulation of CD20 expression during the activation of LN T cells, we examined LNs derived from monkeys experimentally inoculated with simian immunodeficiency virus (SIV). Significant expression of CD20 antigen was detected in the T cells of the LNs at the stage of lymphadenopathy. These findings suggest that lymphocyte activation in the LNs induced expression of the CD20 molecule in some T cells.

Six mouse monoclonal antibodies were developed by immunization with a Japanese monkey cell line. These monoclonal antibodies, designated the UH series, reacted with large populations of peripheral B cells and monocytes, but not with T cells. The distribution of reactivities and the molecular weight of the membrane antigens recognized were similar to those of the HLA-DR monoclonal antibody; one inhibited the binding of HLA-DR. Human interferon-gamma induced increased expressions of all the UH antigen epitopes.

Responses of peripheral blood mononuclear cells to phytohemagglutinin-P (PHA-P), concanavalin-A (ConA), and pokeweed mitogen (PWM) were compared in man and Japanese monkeys. Both CD8+ and CD8- T subsets showed greater responses to ConA than to PHA-P in the Japanese monkey. Addition of macrophages to each T subset produced more effective augmentation of ConA response in the Japanese monkey than in man, and ConA induced more interleukin-2-receptor-positive blast cells than PHA-P did in Japanese monkeys.

A cell line (JAMH17+) resistant to 8-azaguanine was established from a human T-cell leukemia virus type 1 related virus (simian T-cell leukemia virus-1) positive Japanese macaque cell line. Lymphoblastic cell lines were established from the peripheral blood mononuclear cells of humans, hominoids, and several species of macaques by coculture with JAMH17+ in hypoxanthine-aminopterin-thymidine medium. HTLV-1 specific antigen was detected in some of the established cell lines. Phenotypic analysis showed that several cell lines of crab-eating macaques expressed Leu11a antigen, which is a marker of human natural killer cells.

Two complement-related activities were found in the haemolymph of the silkworm, Bombyx mori. One was detected as a ‘classical C3-like’ activity (CC3LA) to cooperate with mammalian or anuran ‘classical R3’ reagent to induce enhanced lysis of rabbit IgM-rich haemolysin-sensitized sheep erythrocytes. Another is detected as an alternative pathway inhibiting activity (APIA). Silkworm haemolymph (SWHL) was fractioned by DEAE cellulose, CM cellulose and Sephadex G-200 chromatographies. In every step of the fractionation procedure, these two activities were eluted in the same regions, suggesting that these two activities are carried on a single factor.

Six monoclonal antibodies to Japanese monkey leukocytes were developed. These monoclonal antibodies, designated the U series, cover most kinds of leukocytes (pan T cells, CD8+ cells, CD8+ subset and granulocytes, CD16+ cells, monocytes/macrophages), and should be useful in the immunological analysis of primate models, such as tissue transplants and virus-related diseases, in particular, acquired immune deficiency syndrome (AIDS).

The expression of the epitope detected by the monoclonal antibody Leu-5 and the rosette formation with sheep red blood cells was examined for the mononuclear cells of 25 primate species including human subject. The plot of rosette-forming cells against Leu-5-positive cells enabled us to classify these species into three large groups: hominoids--Old World monkeys (Leu-5-positive cells greater than or equal to rosette-forming cells), New World monkeys (Leu-5-positive cells less than rosette-forming cells), and prosimians (Leu-5-positive cells less than 5%, some of them with both Leu-5-positive cells and rosette-forming cells less than 5%).

The CD16+ lymphoblastic cell lines of crab-eating monkeys shared the U-5 antigen recognized by a monoclonal antibody. The CD16+U-5+ cell lines expressed high natural killer activity to K562 cells, whereas the CD16-U-5- control cell line had no significant natural killer activity. A possible involvement of the U-5 antigen in natural killer function was also suggested by reduction of the natural killer activity in peripheral blood mononuclear cells of Japanese monkeys after treatment with U-5 monoclonal antibody and complement.

Serum of the Japanese smooth dogfish, Triakis scyllia , was nonhemolytic against rabbit red blood cells (RRBC) and sheep red blood cells (SRBC), but enhanced the activities of heterologous fish sera (sera from Orectolobus japonicus , Squalus mitsukurii and the carp, Cyprinus carpio ) to lyse red cells (RRBC or SRBC) except in case of RRBC- lysis by carp serum. Addition of sub-hemolytic concentration of Squalus or carp serum induced SRBC-lysing activity of Triakis serum. The reaction involved in this enhancement of RRBC (or SRBC)-lysis required divalent cations, Ca ++ and/or Mg ++ , and therefore a complement-like and direct red cell membrane attack by Triakis serum was strongly suggested. Zymosan-treated carp serum did not cooperate with Triakis serum to generate enhanced SRBC-lysis, which indicated a cooperative reaction between Triakis serum factor(s) and carp late complement component(s). Zymosan-treatment reduced the hemolytic activity of the carp serum, whereas it did not cause any detectable reduction in the hemolytic activity of Squalus and Orectolobus sera.

Tissues, such as the fat body, susceptible to a nuclear-polyhedrosis virus developed a sequential synthesis of viral DNA, viral messenger RNA, and viral protein almost 50 hr after virus inoculation, and, thereafter, synthesized polyhedron protein until the late period of infection. On the other hand, in such organs as the gut and the posterior part of the silk gland, which resisted virus infection, DNA was synthesized at the beginning of infection, but afterwards, the synthesis of RNA and protein, that were specific for the virus, was insufficient to adversely affect the function of the cell. It was evident from the data pertaining to the incorporation rate of the precursor of each macromolecule that the fat body could synthesize most of the DNA, RNA, and protein required for virus multiplication in a larva. Analysis of the fat body, and consideration of the rate of incorporation of precursor and the amount of the internal pool, showed that the amount of DNA synthesized in the first several hours after virus inoculation was nearly equivalent to the virus titer 5 of LD 50 , and also, the amount of virus protein synthesized between 7 and 50 hr after virus inoculation was close to 5 of LD 50 . The functional alteration was partly visible at the very beginning of the infection as shown by the fibroin synthesis and the “degeneration” of the nucleus supposedly caused by the remarkable synthesis of DNA in the infected cell.

Publisher Summary Lamprey hemolytic system appears very different from gnathostome complement system. This chapter discusses a study to examine the properdin-like pathway in the serum of the lamprey (Lampetra japonica). Lamprey sera (LS) reacted with rabbit red cells (RRBC) through two successive steps. RRBC-lysis through these complements was inhibited by pretreatment of RRBC with LS, whereas no such Inhibition was observed for LS-treated EA. This difference between the results of RRBC and EA strongly suggest that some site(s) on RRBC capable of reacting with properdin system might be involved in the interaction between RRBC and LS. In the first step, factor X in LS reacted with RRBC, in the absence of divalent cations, to generate an intermediate complex, RRBC-X. In the second step, RRBC-X reacted with some other factor(s) in LS to further generate RRBC, and this step was Mg2+-dependent. Both RRBC and RRBC-X activated, but RRBC did not activate bullfrug complement in EGTA.GVB.2Mg2+. These results strongly suggested a primitive properdin or properdin-like system in the lamprey.

Publisher Summary Mammalian complement (Cf) receptors control the fate of immune complexes and the triggering of the immune response. This chapter discusses the evolution of the vertebrate complement system and immune adherence reaction. It discusses a study to examine whether C3b–R on huE react with various heterologus vertebrate complements. One drop of sample fresh serum from varying vertebrates was 2-fold serially diluted in gelatin veronal buffered saline containing Ca++ and Mg++ by using microtitration plate and added with 1 drop of the suspension of rabbit IgM-rich hemolysin-sensitized sheep erythrocytes (EA) or unsensitized ones (E). After 1 hr incubation of the mixtures, C' titers (CH50) were determined by reciprocals of serum dilution giving 50% lysis of EA or E.