scholarly journals IMMUNOLOGICAL MEMORY IN VITRO

1971 ◽  
Vol 133 (4) ◽  
pp. 846-856 ◽  
Author(s):  
Gordon N. Radcliffe ◽  
Michael A. Axelrad
Keyword(s):  
Spleen Cell ◽  
Primary Response ◽  
Secondary Response ◽  
Spleen Cells ◽  
Sheep Erythrocytes ◽  
Early Primary ◽  
Primary Type ◽  
Type Response

The immune responses to sheep erythrocytes of mouse spleen cell suspensions from immune and nonimmune donors were compared in vitro. In vivo immunity was only transiently reflected in vitro, and 8 wk after in vivo immunization the responses of cultures from immunized and nonimmunized mice were virtually identical. There appeared to be two mechanisms for an antibody response to sheep erythrocytes. The first was responsible for the early primary response and is unmodified in the immune animal though contributing little to subsequent in vivo responses due to its suppressibility by specific antibody. The second was expressed in the in vivo secondary response but not on in vitro challenge of spleen cells from mice immunized many weeks previously; spleen cell cultures from such immune mice, freed from the antibody of the in vivo environment, once again demonstrate a pure primary-type response.

AGGLUTININ RESPONSE TO SHEEP ERYTHROCYTES IN MICE FOLLOWING INTRAPERITONEAL INJECTION OF ALLOGENEIC SPLEEN CELLS

1963 ◽  
Vol 41 (1) ◽  
pp. 573-578
Author(s):  
Gordon O. Bain
Keyword(s):  
Spleen Cell ◽  
Intraperitoneal Injection ◽  
Depressant Effect ◽  
Spleen Cells ◽  
Sheep Erythrocytes ◽  
C3h Mice

C3H mice were injected with C57L spleen cells and 24 hours later with sheep erythrocytes. Recipients of spleen cells from C57L donors previously injected with C3H tissue developed lower antisheep hemagglutinin titers than control mice. Neither donor serum nor saline extracts of lyophilized donor spleen cells had significant cytotoxicity demonstrable in vitro. However, both heat-killed spleen cells and lyophilized spleen cells injected in vivo had a depressant effect on the anti-sheep hemagglutinin titer. The depressant effect is attributed to isoimmunization of the spleen cell donors since injection of spleen cells from donors not previously exposed to C3H antigens did not result in a lowered antisheep hemagglutinin titer.

AGGLUTININ RESPONSE TO SHEEP ERYTHROCYTES IN MICE FOLLOWING INTRAPERITONEAL INJECTION OF ALLOGENEIC SPLEEN CELLS

1963 ◽  
Vol 41 (3) ◽  
pp. 573-578
Author(s):  
Gordon O. Bain
Keyword(s):  
Spleen Cell ◽  
Intraperitoneal Injection ◽  
Depressant Effect ◽  
Spleen Cells ◽  
Sheep Erythrocytes ◽  
C3h Mice

C3H mice were injected with C57L spleen cells and 24 hours later with sheep erythrocytes. Recipients of spleen cells from C57L donors previously injected with C3H tissue developed lower antisheep hemagglutinin titers than control mice. Neither donor serum nor saline extracts of lyophilized donor spleen cells had significant cytotoxicity demonstrable in vitro. However, both heat-killed spleen cells and lyophilized spleen cells injected in vivo had a depressant effect on the anti-sheep hemagglutinin titer. The depressant effect is attributed to isoimmunization of the spleen cell donors since injection of spleen cells from donors not previously exposed to C3H antigens did not result in a lowered antisheep hemagglutinin titer.

scholarly journals IMMUNOLOGIC REACTIONS TO HAPTENS ON AUTOLOGOUS CARRIERS

1973 ◽  
Vol 137 (2) ◽  
pp. 411-423 ◽  
Author(s):  
John W. Moorhead ◽  
Curla S. Walters ◽  
Henry N. Claman
Keyword(s):  
T Cells ◽  
B Cells ◽  
Aminocaproic Acid ◽  
Single Amino Acid ◽  
Secondary Response ◽  
Spleen Cells ◽  
Activated T Cells ◽  
Thymus Cells

Both thymus-derived (T) and bone marrow-derived (B) lymphocytes participate in the response to a hapten 4-hydroxy-3-iodo-5-nitrophenylacetic acid (NIP), coupled to a nonimmunogenic isologous carrier, mouse gamma globulin (MGG). Spleen cells from mice immunized with NIP-MGG show increased DNA synthesis in vitro when cultured with NIP-MGG. The participation of and requirement for T cells in the response was demonstrated by treating the spleen cells with anti-θ serum. This treatment resulted in a 77% inhibition of the antigen response. Furthermore, adoptively transferred normal thymus cells could be specifically "activated" by NIP-MGG in vivo and they responded secondarily to the antigen in vitro. The active participation of B cells in the secondary response was demonstrated by passing the immune spleen cells through a column coated with polyvalent anti-MGG serum. Column filtration reduced the number of NIP-specific plaque-forming cells and NIP-specific rosette-forming cells (both functions of B cells) and produced a 47% inhibition of the NIP-MGG response. The ability of the cells to respond to phytohemagglutinin (PHA) was not affected by column filtration showing that T cells were not being selectively removed. The participation of B cells in the in vitro NIP-MGG response was also shown by treatment of the spleen cells with antiserum specific for MGG and MGG determinants. B cells were removed by treatment with anti-IgM or polyvalent anti-MGG serum plus complement, resulting in a respective 46 and 49% inhibition of the response to NIP-MGG. (Treatment with anti-IgM serum had no effect on T cells.) The contribution of the hapten NIP to stimulation of T cells was investigated using NIP-MGG-activated thymus cells. These activated T cells responded in vitro very well to the NIP-MGG complex but not to the MGG carrier alone demonstrating the requirement of the hapten for T cell stimulation. The response was also partially inhibited (41%) by incubating the activated cells with NIP coupled to a single amino acid (epsilon-aminocaproic acid) before addition of NIP-MGG. These results demonstrated that T cells recognize the hapten NIP when it is coupled to the isologous carrier MGG.

Parasitology and immunology of mice vaccinated with irradiated Litomosoides sigmodontis larvae

Parasitology ◽  
2000 ◽  
Vol 120 (3) ◽  
pp. 271-280 ◽  
Author(s):  
L. LE GOFF ◽  
C. MARTIN ◽  
I. P. OSWALD ◽  
P. N. VUONG ◽  
G. PETIT ◽  
...  
Keyword(s):  
Immune Responses ◽  
Recovery Rate ◽  
Spleen Cells ◽  
Challenge Inoculation ◽  
Infective Larvae ◽  
Litomosoides Sigmodontis ◽  
Before And After ◽  
Type Response

This study was performed with Litomosoides sigmodontis, the only filarial species which can develop from the infective larvae to the patent phase in immunocompetent laboratory BALB/c mice. Parasitological features and immune responses were analysed up to 3 months before and after challenge inoculation, by comparing 4 groups of mice: vaccinated challenged, challenged only, vaccinated only, and naive mice. Male larvae were very susceptible to irradiation and only female irradiated larvae survived in vivo. Protection, assessed by a lower recovery rate, was confirmed and was established within the first 2 days of challenge. This early reduction of the recovery rate in vaccinated challenged mice was determined by their immune status prior to the challenge inoculation. This was characterized by high specific IgM and IgG subclass (IgG1, IgG2a and IgG3) levels, high specific IL-5 secretion from spleen cells in vitro and a high density of eosinophils in the subcutaneous connective tissue. Six h after the challenge inoculation, most tissue eosinophils were degranulated in vaccinated challenged mice. Thus, in the protocol of vaccination described, protection appeared mainly to result from the stimulation of a Th2 type response and eosinophils seemed to be the main effectors for the increased killing of infective larvae in vaccinated challenged mice. Two months after challenge inoculation, the percentage of microfilaraemic mice was lower in vaccinated challenged mice as a consequence of this overall reduction in the worm load. In both vaccinated challenged and challenged only groups, the in vitro splenocyte proliferative capacity was reduced in microfilaraemic mice.

scholarly journals THE EFFECTS OF ANTIGENS AND OF PHYTOHEMAGGLUTININ ON RABBIT SPLEEN CELL SUSPENSIONS

1966 ◽  
Vol 124 (4) ◽  
pp. 621-634 ◽  
Author(s):  
G. Harris ◽  
R. J. Littleton
Keyword(s):  
Dna Synthesis ◽  
Spleen Cell ◽  
Cell Suspensions ◽  
Red Cells ◽  
Specific Response ◽  
Spleen Cells ◽  
Sheep Red Cells ◽  
Stimulation Of

Phytohemagglutinin (PHA) stimulated the rate of DNA synthesis in rabbit spleen cell suspensions. Unlike antigens, previous immunization to PHA was not necessary and the specific response could not be transferred by macrophages, although lymphocytes primed by incubation in PHA were able to stimulate other spleen cells not directly exposed to PHA. When rabbits were stimulated by in vivo immunization with antigens, spleen cells proliferating in response to antigen were stimulated to divide by in vitro contact with PHA. Using the technique of specific hemolytic plaque formation by individual cells synthesizing γM-antibody to sheep red cells (plaque-forming cells), no evidence was obtained that stimulation of cell division by PHA resulted in specific antibody formation, although the presence of antigen resulted both in stimulation of cell proliferation and the production of plaque-forming cells. The presence of both sheep red cells and PHA in the medium of the same cell suspensions did not enhance the production of plaque-forming cells although there was a summative effect on DNA synthesis.

scholarly journals Murine syngeneic mixed lymphocyte response. I. Target antigens are self Ia molecules

1981 ◽  
Vol 154 (5) ◽  
pp. 1652-1670 ◽  
Author(s):  
LH Glimcher ◽  
DL Longo ◽  
I Green ◽  
RH Schwartz
Keyword(s):  
T Cells ◽  
Culture Conditions ◽  
Primary Response ◽  
Secondary Response ◽  
Mouse Serum ◽  
Lymphocyte Response ◽  
Radiation Induced ◽  
Antigen Presenting

A system has been described that produces a murine syngeneic mixed lymphocyte response (MLR) comparable in magnitude to an allogeneic MLR. The responder cells in these cultures exhibit the classic immunologic characteristics of both memory and specificity. Studies using radiation-induced bone marrow chimeras of F(1) {arrow} parent type indicated that, similar to many other T cell-mediated immune responses, the response of the T lymphocytes in the syngeneic MLR was major histocompatibility complex-restricted and was determined by the environment in which the T cells matured. Using responder T cells from F(1) {arrow} parent chimeras and stimulator cells from H-2 recombinant strains, it was possible to map the genes involved in the stimulation to the K and/or I regions. In addition, blocking studies with monoclonal anti-Ia antibodies suggested that in the B10.A strain the critical molecules were products of both the I-A(k) and I-E(k) subregions. The issue of whether the syngeneic MLR is directed solely at self I-region antigens or whether the response represents proliferation to an unknown antigen in association with self I-region determinants was also addressed. Secondary syngeneic MLR were successfully performed in normal mouse serum and with stimulator cells prepared in the absence of bovine serum albumin to rule out the possibility that xenogeneic serum antigens were involved in the stimulation. The possibility that the syngeneic MLR might represent a secondary response to environmental antigens was eliminated by using germ- free mice as a source of stimulator cells and by demonstrating that spleen cells from unimmunized, fully allogeneic chimeras (B10.A {arrow} B10) could generate a normal syngeneic MLR even though such chimeras could not be primed to respond to any foreign antigens unless supplemented in vivo with a source of antigen-presenting cells syngeneic to the B10 host. The possibility that the syngeneic MLR was a primary response to a foreign antigen was considered unlikely because by using our culture conditions we could not obtain a primary antigen response or a secondary antigen response after in vitro priming to a variety of potent foreign antigens. Finally, the possibility that the syngeneic MLR represents a response to a variety of minor histocompatibility self antigens in association with self Ia molecules was eliminated by showing that the secondary responses to H-2 compatible, non-H-2 different strain (A/J vs. B10.A and C3H, or BALB/c vs. B10.D2 and DBA/2) were comparable to the secondary responses to syngeneic stimulators. Thus, we conclude that the target antigens in the syngeneic MLR are solely determinants on self Ia molecules, although the functionally equivalent possibility of a single, nonpolymorphic, minor self antigen seen in association with self Ia molecules cannot be excluded.

scholarly journals THE PROLIFERATIVE AND ANAMNESTIC ANTIBODY RESPONSE OF RABBIT LYMPHOID CELLS IN VITRO

1970 ◽  
Vol 131 (5) ◽  
pp. 970-980 ◽  
Author(s):  
G. A. Theis ◽  
G. J. Thorbecke
Keyword(s):  
Cell Interaction ◽  
Lymphoid Cells ◽  
Secondary Response ◽  
Adherent Cell ◽  
Spleen Cells ◽  
Sheep Erythrocytes ◽  
Absorbent Cotton ◽  
Depletion Effect ◽  
Made In

Both primary and secondary responses to sheep erythrocytes and to Brucella abortus antigen have been obtained in cultures of dispersed rabbit spleen cells. Removal of adherent cells by repeated incubation of spleen cells on absorbent cotton diminished the ability of the spleen cell suspensions to give secondary as well as primary responses in vitro. When comparing cultures made in dishes and in tubes, the loss of responsiveness after incubation on cotton was much more evident in the dish cultures. It was concluded that the cell-to-cell interaction needed for immune responses to particulate antigens in vitro was more readily interfered with when the cells were spread over a larger surface area. The proliferative response to antigen, as measured by uptake of 3H-thymidine in tube cultures of the sensitive spleen cells, appeared particularly resistant to the depletion effect of adherent cell removal. Dispersed spleen cells from sensitized mice gave a secondary response to sheep erythrocytes. This response was readily abolished by one incubation on absorbent cotton when the cells were cultured in dishes.

scholarly journals CD4pos, NK1.1pos T cells promptly produce interleukin 4 in response to in vivo challenge with anti-CD3.

1994 ◽  
Vol 179 (4) ◽  
pp. 1285-1295 ◽  
Author(s):  
T Yoshimoto ◽  
W E Paul
Keyword(s):  
T Cells ◽  
Spleen Cell ◽  
Great Majority ◽  
Cell Suspensions ◽  
Interleukin 4 ◽  
Cellular Interactions ◽  
Spleen Cells ◽  
Ifn Gamma

Injection of anti-CD3 antibodies causes prompt expression of interleukin (IL)-4, IL-2, and interferon gamma (IFN-gamma) mRNA among spleen cells. The optimal dose of anti-CD3 for such induction was 1.33 microgram/animal; lymphokine mRNA was first observed at 30 min, peaked at 90 min, and was undetectable (for IL-4) or had declined markedly by 4 h. Cells harvested from spleens of mice injected with anti-CD3 90 min earlier secreted IL-4, IL-2, and IFN-gamma without further stimulation. By contrast, in vitro stimulation with anti-CD3 of spleen cell suspensions or splenic fragments from noninjected donors failed to cause prompt production of IL-4 and, even after 24 h of stimulation, the amount of IL-4 produced in such cells was substantially less than that secreted within 1 h by spleen cell suspensions or splenic fragments from mice injected with anti-CD3 90 min earlier. Production of IL-4 by spleen cells from anti-CD3-injected mice was not inhibited by pretreatment with anti-IL-4 antibody or with IFN-gamma or tumor growth factor beta nor enhanced by treatment with IL-4. By contrast, CTLA-4 immunoglobulin (Ig) treatment clearly diminished IL-4 production in response to in vivo anti-CD3, indicating that cellular interactions involving CD28 (or related molecules) were important in stimulation. Cell sorting analysis indicated that the cells that produced IL-4 in response to in vivo injection of anti-CD3 were highly enriched in CD4pos cells with the phenotype leukocyte cell adhesion molecule-1 (LECAM-1)dull, CD44bright, CD45RBdull, NK1.1pos. Indeed, the small population of CD4pos, NK1.1pos cells had the great majority of the IL-4-producing activity of this population. Injection with Staphylococcal enterotoxin B also caused prompt induction of IL-4 mRNA; the cells that were principally responsible for production also had the phenotype of CD4pos, NK1.1pos. These results suggest that possibility that this rare population of T cells may be capable of secreting IL-4 at the outset of immune responses and thus may act to regulate the pattern of priming of naive T cells, by providing a source of IL-4 to favor the development of T cell helper 2-like IL-4-producing cells.

scholarly journals GAMMA GLOBULIN AND ANTIBODY FORMATION IN VITRO

1967 ◽  
Vol 125 (1) ◽  
pp. 33-44 ◽  
Author(s):  
Vera J. Stecher ◽  
G. Jeanette Thorbecke
Keyword(s):  
Antibody Formation ◽  
Primary Response ◽  
Secondary Response ◽  
Detectable Effect ◽  
Complete Suppression ◽  
Similar Time ◽  
X Irradiation ◽  
Early Radiation

The present studies have shown that the influence of X-irradiation on the secondary antibody response in vitro is remarkably similar to its effect on the primary response in vivo. When sensitized tissue was first irradiated and then reexposed to antigen, the duration of the interval between irradiation and antigen addition determined the degree of inhibition of the secondary response obtained. A delay of 12 hr resulted in stronger inhibition than a delay of 6 hr, and an interval of 24 hr before reexposure to antigen caused complete suppression of antibody production to diphtheria toxoid and almost complete suppression when sheep RBC were used as the antigen. Induction of the secondary response in rabbit lymph node tissue in vitro followed by exposure to X-irradiation, revealed that immediate exposure to irradiation after antigen produced stronger inhibition of the subsequent response than irradiation on days 2–3. Irradiation on day 6 had no detectable effect. The effectiveness of the early radiation is probably due to prevention of the proliferation of the antibody-forming cells. BUDR was found to be effective at similar time periods as X-irradiation, whereas colchicine could still stop antibody formation when added late during the secondary response in vitro. It was noted that lymph nodes from some BSA-sensitized rabbits as late as 18 months after sensitization gave a response indistinguishable from a typical secondary response, even when not reexposed to antigen.

scholarly journals Duration of the initial TCR stimulus controls the magnitude but not functionality of the CD8+ T cell response

2006 ◽  
Vol 203 (9) ◽  
pp. 2135-2143 ◽  
Author(s):  
Martin Prlic ◽  
Gabriela Hernandez-Hoyos ◽  
Michael J. Bevan
Keyword(s):  
T Cells ◽  
Diphtheria Toxin ◽  
Clonal Expansion ◽  
Primary Response ◽  
T Cell Response ◽  
Secondary Response ◽  
Memory Cells ◽  
Effector Cells

CD8+ T cells only require a brief stimulation with antigen in vitro to divide and differentiate into effector and memory cells upon transfer in vivo. The efficiency of clonal expansion and the functional characteristics of memory cells derived from briefly stimulated cells are poorly defined. We developed a system that allowed us to examine programming entirely in vivo. This was achieved by rapidly killing peptide-pulsed DCs carrying a diphtheria toxin receptor transgene with timed injections of diphtheria toxin without altering the course of an accompanying infection. The magnitude of clonal expansion, but not the functionality of the effector cells, correlated directly with the duration of antigen exposure. Furthermore, memory T cells were capable of mounting a secondary response, regardless of the length of antigen encounter during the primary response. These results indicate that the duration of initial antigen encounter influences the magnitude of the primary response, but does not program responsiveness during the secondary challenge.

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