Immune Lymph Node Cells Research Articles

Intestinal mast cells and their relation to mesenteric lymph nodes.

The data presented in this paper support a common source for intestinal mast cells in intestinal mucosae. IMC precursors and/or mastocytopoietic factors of as yet unknown origin could be transferred by immune mesenteric lymph node (IMLN) cells of thymus-bearing mice. The differentiated locally or induced mastocytopoiesis in Trichinella spiralis infected mucosal sites, but only in animals with a normal thymus function. Antigen-stimulated precursor cells either in the gut or in the mesenteric lymph node to enter the circulation and home to both parasitized and nonparasitized mucosal sites. Both proliferation in the mesenteric lymph node and in the intestinal mucosae were shown to be thymus-dependent. The injected IMLN stimulated only the proliferation of IMC and not of globule leucocytes. This might indicate that cells relevant for IMC proliferation are different from those essential for GL proliferation.

Thymus dependence of the adoptive transfer of intestinal mastocytopoiesis in Trichinella spiralis-infected mice.

The relationship between the gut and mesenteric lymph node in the kinetics of mucosal mast cells (MMC) was further analysed. A Trichinella-induced synchronous proliferation of MMC occurred in the intestine of thymus-bearing and not in athymic mice, both at sites of parasite invasion (jejunum) and elsewhere (caecum) and in the mesenteric lymph node. The mastocytopoietic ability could be adoptively transferred with immune mesenteric lymph node cells to the jejunum and caecum of infected thymus-bearing recipients. The enhanced mastocytopoiesis which was both antigen- and thymus-dependent, was only observed for the stroma intestinal mast cells (IMC) and not for the intraepithelially located globule leucocytes (GL). The data suggest a common source for IMC in intestinal mucosae and support the idea that IMC and GL are two independent cell populations.

Soluble factors in tolerance and contact sensitivity to 2,4-dinitrofluorobenzene in mice. IV. Characterization of migration inhibition factor-producing lymphocytes and genetic requirements for activation.

The production of migration inhibition factor (MIF) in vitro by lymph node cells from mice with contact sensitivity to 2,4-dinitrofluorobenzene (DNFB) was investigated. MIF activity of cell-free culture supernatants was measured using a micro, indirect "hanging-drop" assay system. We found that DNFB-sensitized lymph node cells are stimulated to produce MIF by co-culture with DNP-labeled spleen cells or splenic adherent cells. The stimulation was quantitatively antigen-specific, as co-culture with TNP-spleen cells or TNP-splenic adherent cells induced only low levels of MIF activity. Pretreating the immune lymph node cells with different antisera plus complement, before addition of DNP-spleen cells, showed that MIF production is dependent on Ia- T cells. Additional experiments showed that in order for the T cells to be stimulated, homology at the I-A subregion of the major histocompatibility complex between the T cells and DNP-spleen cells is required. Collectively, these results correlate with our previous finding that transfer of contact sensitivity is mediated by Ia- T cells and indicate that both tests, i.e., transfer in vivo and MIF production in vitro, are measuring effector functions of the same T cell subset.

Immunity to Strongyloides ratti in rats. 1. Adoptive transfer with mesenteric lymph node cells.

Adoptive transfer of immunity against the enteral phase of Strongyloides ratti was monitored using four parameters; namely, number of worms recovered, position occupied in the host's small intestine, worm length and fecundity. When immune mesenteric lymph node cells (IMLNC) were transferred from donors infected for 20, 26 or 32 days into recipients infected for 3 days, a marked acceleration of worm expulsion was evident by day 16 post-infection (p.i.). IMLNC from day-16 p.i. donors did not transfer expulsion. In an experiment in which recipients were given 2 x 10(8) or 1 x 10(8) IMLNC from donors infected for 26 days, accelerated worm expulsion occurred only with the higher inoculum, although manifestations of direct worm immunity, i.e. altered position and reduced length and fecundity, were evident in both cases. Transfer of IMLNC appeared to have no effect upon worm establishment; when cells were transferred on the day of infection a period of 16 days was necessary to effect both direct anti-worm immunity and expulsion, although the former was evident on day 14 p.i. However, when IMLNC were transferred to rats already infected for 6 days, a very significant reduction of their worm burden was apparent 10 days after IMLNC transfer.

Transfer of immunity to Nematospiroides dubius: co-operation between lymphoid cells and antibodies in mediating worm expulsion.

The effect of transferring immune serum (IS) and immune mesenteric lymph node cells (IMLNC) either alone or in combination was studied in NIH mice infected with partially radiation attenuated (5 krad) N. dubius. It was demonstrated that immunity to N. dubius could be transferred with IS and with IMLNC. However, considerably greater protection was transferred to recipient mice when they received both IS + IMLNC. Animals treated in this way had fewer worms than either of the other groups from as early as day 9 onwards, suggesting that a substantial proportion of the worms in mice given IS + IMLNC was retained in the intestinal tissues. The few surviving worms which completed the tissue phase of their development were then rejected by the fourth week of infection. Dose response data showed that as few as 1 x 10(7) IMLNC could cause a significant reduction in worm numbers when given in combination with IS. These experiments indicate that both antibodies (IS) and sensitized lymphoid cells (IMLNC) are required for effective resistance to N. dubius.

Strongyloides ratti: Transfer of immunity in rats by lymph node cells or serum

Immune mesenteric lymph node cells (IMLNC) and hyperimmune serum, alone or in combination, were transferred to recipient, syngeneic Lewis rats. On the day of cell transfer, the recipients, along with appropriate controls, were infected with 1,000Strongyloides ratti larvae. Twelve days later, the number of adult worms in the anterior small intestine was counted and the number ofS. ratti eggs in the terminal uterus of representative female worms was determined. Using these methods, we showed that worm fecundity was reduced in the IMLNC recipients, the hyperimmune serum recipients, and the recipients of both cells and serum as compared with regular controls (averages of 2.45, 2.93, and 2.98 eggs, respectively, vs. 4.73 eggs). The number of adult worms recovered from the IMLNC recipients was not reduced (474.6 worms). However, significantly fewer worms were recovered from the recipients of IMLNC-hyperimmune serum and the recipients of hyperimmune serum only as compared with regular controls (184.8 and 171.5 worms, respectively, vs. 567.2 worms). A working hypothesis, involving an interaction of serum and cellular components, is postulated as the mechanism for this protective response.

Transfer of immunity to Babesia microti of human origin using T lymphocytes in mice

Lymph node and spleen cells from mice infected with Babesia microti of human origin developed the ability to transfer adoptive immunity to naive mice within 25 days after infection. This protective activity was greater in cells obtained at 32 days than in cells obtained at 25 days postinfection and remained stable up to 52 days postinfection. Recipients of lymph node cells and spleen cells displayed similar peak parasitemias although 2 days after peak parasitemia, immune spleen cell recipients had significantly lower parasitemias than immune lymph node cell recipients. Strong protective activity was demonstrated when cells were transferred 1 day postinfection, while equal numbers of cells, transferred 3 days postinfection did not confer significant protection over nonimmune cells. There was also a suggestion that the number of immune spleen cells necessary for significant protection was directly related to the number of parasites inoculated. The subpopulation of lymphocytes responsible for the transfer of adoptive immunity to B. microti of human origin was then studied in BALB/c mice depleted of T lymphocytes by thymectomy and lethal irradiation. One day after infection with B. microti, T-cell-depleted mice were given complement-treated immune spleen cells, anti-θ serum-treated immune spleen cells, nonimmune spleen cells, or no cells. Similar experiments were performed comparing the effects of anti-immunoglobulin serum-treated and unfractionated immune spleen cells on B. microti parasitemia. Treatment with anti-θ serum abrogated the protective activity of immune spleen cells while anti-immunoglobulin serum treatment had no effect. These results suggest that immunologic memory of B. microti in BALB/c mice is modulated by T rather than B lymphocytes.

Murine delayed hypersensitivity (DHS): Modulation of in vitro antigen-induced responsiveness of lymph node cells from mice expressing DHS to human γ-globulin by lymphocyte populations from normal syngeneic animals

Using a chemically defined, protein-free medium, the modulatory effect of normal (N) lymphocytes on in vitro antigen-induced proliferation by lymph node cells (LNC) from mice immunized to express delayed hypersensitivity (DHS) to human γ-glogulin (HGG) was quantitated in coculture. LNC from normal syngeneic animals exerted little if any effect on immune-LNC proliferation. Compared with immune-LNC plus N-LNC coculture response. N thymus cells (TC) were consistently suppressive while N spleen cells (SC) varied in their effect from a marginal to a marked potentiation of radiolabeled thymidine incorporation. Inactivation of N-SC suspensions by X irradiation prior to coculture with immune LNC abrogated the increased responsiveness. It therefore appeared that interaction of immune LNC and antigen resulted in recruitment of N-SC to proliferate. Separation of N-SC suspensions to provide enriched populations of thymic-independent (B) and thymic-dependent (T) lymphocytes showed that B cells augmented and T cells suppressed HGG-induced incorporation of [3H] thymidine when cocultured with immune LNC.

Immunity to Trichinella spiralis in irradiated mice

Irradiation prevented the accelerated expulsion of Trichinella spiralis from mice immunized by transfer of immune mesenteric lymph node cells (IMLNC) or by prior infection. Nevertheless, worms in irradiated immune mice were smaller and less fecund than those in controls. In adoptively immunized and irradiated mice expulsion could not be achieved by increasing the numbers of IMLNC transferred, although the effect upon worm length was more severe. Thus IMLNC express a direct, anti-worm immunity which is independent of their role in worm expulsion. IMLNC cause expulsion in irradiated mice only when adequate levels of bone marrow-derived cells are available. The results are discussed in terms of a possible antibody-mediated basis for direct anti-worm immunity.

Effect of Immunologic Intervention on In Vivo Murine Rous Sarcoma Virus Tumorigenesis2

The effect of immunologic manipulation on the development of primary murine Rous sarcoma was examined after neonatal injection of Rous sarcoma virus-induced chicken tumor material (RCTM) into mice. Heterologous antithymocyte serum (ATS) treatment of mice beginning at 7 and 21 days of age tripled and nearly doubled, respectively, the incidence of primary Rous sarcoma, but treatment at 21 days significantly prolonged the mean latency period of the resultant tumors. Also studied was the effect of an ip injection of syngeneic adult mouse lymphold cells, either normal or sensitized, on primary Rous sarcoma incidence. Lymphold cells from syngeneic adult mice immunized with allogeneic Rous sarcomas (and resistant to syngeneic tumor challenge) had no effect when the animals received infections at birth (day 1), but immune lymph node cells (LNC) significantly increased tumor incidence when animals were given injections of LNC 1 week after birth and injections of RCTM on day 1. LNC and spleen cells (SC) from syngeneic adult mice that had been inoculated with RCTM at birth but had not developed tumors by 200 days of age (V+T−) decreased tumor incidence when administered on day 1 but not on day 7. Normal adult SC and LNC had no effect when given on day 1 but significantly increased tumor incidence when given on day 7. The tumor-stimulating activity of normal lymphoid cells administered on day 7 of life was observed both in the relatively susceptible strain C57BL/10ScSn mice and the resistant H-2 congenic strain B10.D2. Serum from adult immune, V+T−, and T+ mice administered on days 1 and 7 decreased primary tumor incidence, though only the effect of T+ serum was statistically significant. The offspring of immune mothers were less susceptible than those of normal mothers. The offspring of passively immunized mothers given serum from immune females during pregnancy were more susceptible than controls. RCTM-inoculated mice were apparently susceptible only to immunologic manipulations that reduced primary tumor incidence during the first few days of life. After 7 days of age, all treatments had no net effect or increased tumor incidence, though ATS treatment beginning at day 21 significantly lengthened the mean tumor latency period.

A cell-mediated reaction against glomerular-bound immune complexes.

Lewis rats were given a single i.v. injection of soluble immune complexes containing human serum albumin (HSA) and rabbit anti-HSA antibodies, prepared in antigen excess. This resulted in localization of HSA and rabbit gamma globulin (RGG) in glomerular mesangial regions without producing definite histologic changes. 24 h after the injection of immune complexes, groups of these rats received lymph node cells or T-cell preparations from syngeneic donors sensitized to RGG, HSA, or ovalbumin; another group received no cells. All of these groups and a group of normal control rats were given injections of [3H]thymidine at 18, 27, and 44 h. The animals were killed 48 h after the time of cell transfer. In histologic sections, glomerular abnormalities were found only in some of the animals that had received immune complexes and lymph node cells or T-cell populations from donors sensitized to HSA or RGG; the lesions were characterized by focal and segmental increase in cells in mesangial regions. Autoradiographs revealed significantly greater numbers of labeled cells in mesangial regions and glomerular capillaries in the groups that had received immune complexes and cells from HSA- or RGG-sensitized donors than in any of the other groups. Electronmicroscopic studies suggested that the increase in cellularity in mesangial regions resulted from an influx of mononuclear phagocytes. The findings indicate that cell-mediated reactions can be initiated by the interaction between sensitized T lymphocytes and antigens present in immune complexes within mesangial regions.

Soluble factors in tolerance and contact sensitivity to 2,4-dinitrofluorobenzene in mice. III. Histocompatibility antigens associated with the hapten dinitrophenol serve as target molecules on 2,4-dinitrofluorobenzene-immune T cells for soluble suppressor factor.

Previous studies have shown that suppression of 2,4-dinitrofluorobenzene (DNFB) contact sensitivity by soluble suppressor factor (SSF) requires that the donor of immune lymph node (LN) cells and of SSF share either the H-2K and/or H-2D region of the major histocompatibility complex. Thus, target or acceptor molecules for SSF appear to be coded for by genes within the H-2K and H-2D loci. Experiments were done to investigate the nature of these target molecules and to determine what cell types expressed them. It was found that purified lymph node T cells are suppressed by SSF indicating that T cells express the acceptor molecules. Adsorption experiments showed that the only cells capable of adsorbing the suppressor factor are DNFB-immune T cells from donors which share with the factor-producing strain either the H-2K or H-2D locus. This adsorption can be specifically blocked by pretreating the immune LN cells with antibodies directed against H-2K and/or H-2D determinants or against the hapten DNP but not by antibodies against Ia or theta-antigens. Collectively, these results indicate that the target molecules are expressed only by DNFB-immune T cells and are comprised of histocompatibility antigens associated with DNP.

Functional diploidy for the secretion of the a allotypes by the lymphoid cell of heterozygous rabbits

Can a given heterozygous lymphoid cell, for the ‘a’ system of allotypes in the rabbit, secrete immunoglobulin molecules of the two types (functional diploidy) with the same antibody specificity? This problem was investigated by two methods (applied to immunized lymph node or spleen cells): enhancement of plaques of hemolysis by separate action of two anti-allotype antibodies vs the effect of a mixture of antisera, or by micromanipulation. Both methods lead to the same results: many cells manifest functional diploidy. In addition, it was found that only two populations of cells existed: cells secreting only al and cells secreting a1 and a2 or a1 and a3.

Are VH framework antigenic determinants expressed on both rabbit B and T lymphocytes?

AbstractThe extent to which VH framework determinants are represented on T cell receptors is unresolved and controversial. We have studied this question in the rabbit using antia (VH framework) allotype antisera and a goat anti‐rabbit VH antiserum. Flow micro‐ fluorometric analysis of spleen lymphocytes from hyperimmune animals (Freund's adjuvant plus picryl chloride) demonstrated closely similar frequencies of cells stained with the anti‐VH framework antisera compared to anti‐light chain antisera. To search for potentially hidden molecules, additional studies were performed with radioiodinated, detergent‐solubilized plasma membranes derived from immune spleen and lymph node cells. The membranes were analyzed by standard immunoprecipitation techniques. Absorption with anti‐Ig constant region immunoabsorbents removed all radioactivity precipitable by specific anti‐VH framework antisera compared to controls, whereas after absorption with control (normal goat globulin) immunosorbents, VH‐bearing surface IgM and putative surface IgD (presumably of B cell origin) were readily demonstrated by immunoprecipitation. Thus, no evidence for a population of cells or a class of molecule(s) bearing VH in the absence of light or heavy chain constant region determinants was obtained. Although these studies do not support the concept of similar VH framework determinants on T and B cell receptors, they do not exclude a much more limited sharing of determinants or a very minor T cell subpopulation which expresses VH framework that would not be detectable by the methods used in these studies.

Differential Protective Effects of Immune Lymphoid Cells against Transplanted Line Ib Leukemia and Immune Polioencephalomyelitis

Abstract The capacity of immune cells obtained from the major lymphoid compartments to protect C58 mice from transplanted line Ib leukemia, and from an age-dependent autoimmune CNS disease (immune polioencephalomyelitis = IPE) elicited by immunizing old C58 mice with inactivated Ib cells was quantified. Cells used for comparative adoptive protection tests were harvested from the major lymphoid compartments 14 to 15 days after young C58 mice were immunized with inactivated Ib cell preparations. Regression curves were plotted from survival data and the log10PD50 values were determined. Immune spleen (ISC) and peritoneal cells (IPEC) were significantly more protective against transplanted Ib cells than immune lymph node (ILNC), thymic (ITC), and marrow cells (IMC). In contrast, IPEC and IMC were not protective against IPE and ITC were only marginally protective. ILNC afforded significant protection to transplantable leukemia but were only marginally protective to IPE. When ISC were treated with anti-thy 1.2 serum and complement, protection against transplanted leukemia and IPE was reduced > 99%. When donors of immune lymphoid cells were treated with 12.5 mg of cortisone acetate daily for 2 days before lymphoid cells were harvested, protection against transplanted Ib cells by ISC was reduced by approximately 90% whereas protection against IPE was totally eliminated. Considered together, these results indicate that the protective mechanisms to transplantable leukemia and IPE differ significantly in the same indicator mouse strain.

Nature of a humoral factor of bone marrow stimulating antibody production

An active factor doubling the antibody synthesis in the culture of immune lymph node cells was isolated from the supernatant of the cultures of bone marrow cells from intact donors. Gel chromatography was used for this purpose. The stimulant of antibody producers (SAP) was eluated at the site of cytochrome C exit; the factor is heat-stable and has the mol wt of 1300 daltons. SAP production needs RNA and protein synthesis, but does not require DNA synthesis. The radioactive label showed that SAP includes ribonucleotides and amino acids. It is suggested that SAP plays an important role in the regulation of antibody synthesis.

Adoptive immunotherapy and radiotherapy in the treatment of urinary bladder cancer.

Thirty-one patients with invasive transitional cell carcinoma of the urinary bladder were treated either by infusion of tumour immune pig mesenteric lymph node cells into the tumour blood supply, followed 6 weeks later by 5500 rad, or by radiotherapy alone. The patients were paired prior to treatment, using 4 criteria: age (nearest decade), sex, clinical stage (T3 or T4) and histological grade (average or high). The first patient in each pair then received combined treatment and the second radiotherapy alone. Administration of pig cells before radiotherapy offered no advantage to the patient in terms of the incidence of remission or the length of survival after treatment.

Effect of host sex on passive immunity in mice infected with Nematospiroides dubius

Dobson C. & Owen M. E. 1978. Effect of host sex on passive immunity in mice infected with Nematospiroides dubius. International Journal for Parasitology 8: 359–364. Female C 3H but not Quackenbush (Q) mice harboured fewer Nematospiroides dubius than male C 3H and Q mice. Both strains lost worms 21 days after infection. C 3H and Q mice became progressively immune to infection following 4 sequential doses of N. dubius larvae and showed a sex resistance to infection. Hypothymic nu/nu CBA Balb/c mice did not show these effects on N. dubius infection. The reciprocal transfer of male and female immune mesenteric lymph node cells (IMLNC) to syngeneic male and female recipients showed that the female environment enhanced the protective qualities of both male and female IMLNC but the male environment suppressed these effects. Gonadectomized male and female recipients of male and female IMLNC had levels of infection similar to the entire female recipients. Serum from immune female donor mice protected both male and female recipients better than immune serum from male donors, but female mice in each treatment group were better protected than male mice. Immune serum transferred greater levels of protection then IMLNC to recipient mice against N. dubius infections. These data are consistent with the conclusion that the male environment suppresses lymphocyte activity and the production of protective antibodies and additionally may depress the effectiveness of sensitized lymphocytes and antibodies in ejecting N. dubius. On the other hand the female environment does not appear to adversely affect the mobilization of the protective immune response and may enhance immune effector mechanisms in protecting mice against N. dubius infections.

Suppression of rejection of Nippostrongylus brasiliensis in iron and protein deficient rats: effect of syngeneic lymphocyte transfer.

Rejection of Nippostrongylus brasiliensis is impaired in iron and protein deficient rats and this suggests that iron and protein deficiency directly or indirectly suppresses the immune response. The site of the immunological defect in deficient rats was investigated using the technique of cellular transfer of resistance. The functional activity of immune mesenteric lymph node cells obtained from iron and protein deficient donors was not depressed as measured by their capacity to cause parasite rejection in nutritionally sufficient recipients. In contrast, immune lymph node cells obtained from either sufficient or deficient donors did not result in parasite rejection in iron and protein deficient recipients. These results indicate that there is no permanent defect of lymphocyte function in iron and protein deficient rats and suggest that either some other component of the rejection mechanism is defective, or that lymphocyte function is blocked in an iron and protein deficient environment.

Comparative Study of Cytotoxic T‐Lymphocytes and Producers of the Macrophage Migration Inhibition Factor (MIF) in the H‐2 System

Comparative study of two T‐cell subpopulations, killers and MIF producers, immune to H‐2 antigens, was performed using the in vitro micro‐techniques of cell‐mediated cytolysis and inhibition of macrophage migration. T‐dependence of MIF production was shown in this system by lymphocyte separation onto T‐ and B‐cells and by inactivation of MIF producers with anti‐θ serum. At least part of MIF producers appeared to be more sensitive than killers to the action of anti‐κ antibodies in the presence of complement. Production of MIF as compared to the cytotoxic activity was revealed in regional lymph node cells earlier and preserved longer after immunization with allogeneic tumour cells. Immune lymph node cells were shown to produce MIF when incubated with spleen cells of congenic and recombinant strains of mice possessing either private or public H‐2 specificities of the immunizing complex, but they did not produce MIF when incubated with syngeneic spleen cells. Removal of killers by absorption on the relevant target cells did not reduce the ability of non‐adherent lymphocytes to produce MIF. Unlike cytotoxic activity, MIF production is inhibited by pre‐treatment of immune lymphocytes with pronase. The data obtained are indicative of essential differences in effector T‐cell subpopulations which exert the cytotoxic effect and produce MIF in the H‐2 system. The implications of these data for studies on T‐cell differentiation under the action of H‐2 antigens are discussed.