Syngeneic transplantation with peripheral blood mononuclear cells collected after the administration of recombinant human granulocyte colony-stimulating factor

Five syngeneic transplants were performed in four patients following myeloablative therapy using unmodified peripheral blood mononuclear cells (PBMCs) collected after the administration of recombinant human granulocyte colony stimulating factor (rhG-CSF) to normal donors. The only toxicity experienced by the four normal donors was bone pain. Four patients received two collections of PBMCs, and a second transplant was performed in one patient with one collection. The patients received a median of 20.53 x 10(8) total nucleated cells/kg (range 20 to 25.5), 11.3 x 10(8) total mononuclear cells/kg (range 6.52 to 17.2), 113.1 x 10(4)/kg CFU-GM (range 46.7 to 211.8) and 9.6 x 10(6) CD34+ cells/kg (range 1.6 to 12.6) Post-transplant growth factors were not administered. The median time to an absolute neutrophil count greater than 0.5 x 10(9)/L was 14 days (range 10 to 18). The median time to platelet transfusion independence was 11 days (range 10 to 13). Two patients had the number of CD3+ T lymphocytes determined in the pheresis product. An average of 3.04 x 10(10) CD3+ cells were collected per pheresis. This represents an approximate 1 log increase over the number of T lymphocytes in a typical bone marrow transplant. Rh-GCSF can be used to mobilize peripheral blood progenitor cells from normal donors with minimal toxicity. Studies of allogeneic transplants using PBMCs collected after rhG-CSF administration to determine permanent grafting ability and the incidence and severity of graft-versus-host disease are warranted.     

Functional T cells in athymic nude mice.

After passage of spleen cells from nu/nu mice over a nylon wool column, concanavalin A-responsive cells can be detected in the presence of 2-mercaptoethanol, and specific cytotoxic T lymphocytes can be generated without exposure to interleukin 2 (IL-2). The spleen cells of the nu/nu mice born of nu/nu parents and nursed by nu/nu mothers had significantly fewer Thy-1+ T cells and a lesser capacity to generate cytotoxic T lymphocytes than did the conventionally bred nu/nu mice. Nonetheless, such cells were clearly present. IL-2 may act to cause these post-thymic T cells to proliferate. Therefore, it seems inappropriate to consider IL-2 as an inducer of the differentiation of T cells in the absence of thymic influence on the basis of the capacity of IL-2 to induce the appearance of a T-lymphocyte population in nu/nu mice.     

A human soluble suppressor factor affecting lymphocyte responses in vitro.

A soluble suppressor factor (SSF) has been demonstrated in the supernatant of normal human peripheral blood lymphocyte cultures that exhibits suppressive activity toward the proliferative response of normal lymphocytes to concanavalin A or alloantigens in mixed lymphocyte culture (MLC) or toward pokeweed mitogen-stimulated immunoglobulin synthesis and secretion in vitro. Suppression of the proliferative response in MLC reached maximal levels when added SSF-containing supernatant approximated 20% by volume of the culture medium. Suppression in the MLC was found to act at the proliferative stage. SSF acts independently of cytotoxicity and is stable at 56 degrees C for 30 min but is inactivated at higher temperatures. Addition of SSF to the MLC as late as day 4 after initiation of the culture results in suppression of transformation. This factor(s) may regulate the magnitude of several immune responses in humans.     

Selective growth of a population of human basophil cells in vitro.

An initially homogeneous population of basophilic polymorphonuclear leukocytes was derived from human fetal liver cells grown in culture for 5-7 days. The cells were characterized as basophils by their morphology, histologic staining characteristics, and histamine content, and by the presence of IgE receptors on their surface. In this culture system the basophils were viable for up to 10 days.     

Growth of a pure population of mouse mast cells in vitro with conditioned medium derived from concanavalin A-stimulated splenocytes.

A pure population of mast cells was obtained after 14 days of culturing mouse bone marrow cells in the presence of medium derived from concanavalin A-stimulated mouse spleen cells. The cells were characterized as mast cells by their morphologic appearance and histologic staining, by their histamine content (450 ng per 10(6) cells) and by the demonstration of IgE receptors on their surface (150,000--440,000 receptor sites per cell). The histamine content and the number of IgE receptors remained constant for at least 7 wk of culture. These mast cells could be passively sensitized to mice hybridoma IgE. They then released 43% of their histamine content upon incubation with anti-mouse hybridoma IgE.     

Influence of dietary restriction on immunologic function and renal disease in (NZB × NZW) F1 mice

In (NZB x NZW)F(1) (B/W) mice, moderate caloric intake [10 kcal (41.8 kJ) per day] from the time of weaning was associated with maintenance of lower body weight, greater capacity of spleen cells to be stimulated with T-cell mitogens, and better preserved capacity to generate cytotoxic cells in response to in vitro and in vivo stimulation with allogeneic tumor cells. Plaque-forming cell response to sheep erythrocytes was also well maintained in animals on the restricted diets when sensitization was accomplished either in vitro or in vivo. Spontaneous suppressor cell activity against plaque-forming cells that developed in controls did not appear in the mice on the restricted diet. Significantly less circulating antibody to native DNA was present in the blood of mice 10 months of age when their dietary intake had been restricted. Histological analysis revealed that the development of renal disease and the deposition of gamma globulin in the glomerular capillaries was markedly inhibited in the mice on restricted diets. Dietary restriction from the time of weaning thus appears to prolong significantly the life of autoimmunity-prone (NZB x NZW)F(1) male and female mice and to alter lymphoid cell immune function, thereby decreasing the autoimmune processes and immunological assault associated with progressive renal disease in these animals.     

Influence of dietary energy restriction on the numbers and proportions of Ly-1+ B lymphocytes in autoimmunity-prone mice.

Chronic energy-intake restriction (CEIR) has been shown to increase life-span, delay diseases expression, and inhibit immunological perturbations in all strains of autoimmunity-prone mice studied, including NZB, (NZB x NZW)F1, MRL/Mp-lpr/lpr, BXSB, and kd/kd mice. In (NZB x NZB)F1 mice, increased percentages and increased absolute numbers of Ly-1+ B lymphocytes in spleen, mesenteric lymph nodes, thymus, and bone marrow as revealed by two-color immunofluorescence analysis were greatly reduced by CEIR with a diet high in carbohydrate and low in fat. This influence on a possibly crucial lymphocyte subpopulation was associated with delayed onset of disease and with greatly prolonged life-span. In the present investigation, the percentages and absolute number of Ly-1+ B lymphocytes in the spleen, peritoneal exudate, and peripheral blood were found to be increased in each autoimmunity-prone strain studied. CEIR decreased the absolute and relative numbers of the Ly-1+ B lymphocytes in mice of each of the autoimmunity-prone strains and returned the number and proportions of Ly-1+ B cells close to levels present in the same locations in genetically long-lived C57BL/6, DBA/2, or BALB/c mice fed a standard commercial diet ad libitum.     

The purine salvage enzyme hypoxanthine guanine xanthine phosphoribosyl transferase is a major target antigen for cell-mediated immunity to malaria

Although there is good evidence that immunity to the blood stages of malaria parasites can be mediated by different effector components of the adaptive immune system, target antigens for a principal component, effector CD4(+) T cells, have never been defined. We generated CD4(+) T cell lines to fractions of native antigens from the blood stages of the rodent parasite, Plasmodium yoelii, and identified fraction-specific T cells that had a Th1 phenotype (producing IL-2, IFN-gamma, and tumor necrosis factor-alpha, but not IL-4, after antigenic stimulation). These T cells could inhibit parasite growth in recipient severe combined immunodeficient mice. N-terminal sequencing of the fraction showed identity with hypoxanthine guanine xanthine phosphoribosyl transferase (HGXPRT). Recombinant HGXPRT from the human malaria parasite, Plasmodium falciparum, activated the T cells in vitro, and immunization of normal mice with recombinant HGXPRT reduced parasite growth rates in all mice after challenge.