scholarly journals MACROPHAGE-LYMPHOCYTE CLUSTERS IN THE IMMUNE RESPONSE TO SOLUBLE PROTEIN ANTIGEN IN VITRO

1974 ◽  
Vol 140 (5) ◽  
pp. 1245-1259 ◽  
Author(s):  
Ole Werdelin ◽  
Otto Brændstrup ◽  
Eskild Pedersen
Keyword(s):  
Immune Response ◽  
Lymph Node ◽  
Cluster Formation ◽  
Peritoneal Macrophages ◽  
Physical Interaction ◽  
Antigen Binding ◽  
Purified Protein Derivative ◽  
Cell Clusters ◽  
Immune Lymphocytes

We have studied the physical interaction between macrophages and lymphocytes during the immune response to purified protein derivative of tuberculin (PPD) in vitro. Mixtures of peritoneal macrophages and lymph node lymphocytes from guinea pigs immunized with tubercle bacilli formed cell clusters during 20 h of culture with PPD. The number of clusters produced was correlated to the number of immune lymphocytes in the cultures. Peritoneal macrophages which had been pulsed with PPD and untreated lymph node lymphocytes produced cell clusters in the absence of free PPD in numbers equivalent to those produced by the same cells in the presence of free PPD. In cultures containing a mixture of PPD-pulsed macrophages, not-pulsed macrophages, and immune lymphocytes with no free PPD, cell clusters developed mainly between the antigen-pulsed macrophages and lymphocytes. Cluster formation was antigen-specific with the specificity residing in the lymphocytes, mainly or exclusively in the T lymphocytes. These data indicate that in the process of cell cluster formation macrophages serve as antigen-binding (or -processing) cells, while a subpopulation of lymphocytes interact physically and specifically with the macrophages.

scholarly journals MATURATION OF THE IMMUNE RESPONSE IN VITRO

1972 ◽  
Vol 136 (2) ◽  
pp. 353-368 ◽  
Author(s):  
Alberto J. L. Macario ◽  
Everly Conway de Macario ◽  
Claudio Franceschi ◽  
Franco Celada
Keyword(s):  
Escherichia Coli ◽  
Immune Response ◽  
Lymph Node ◽  
Association Constant ◽  
Secondary Response ◽  
Antibody Specificity ◽  
Affinity Selection ◽  
Immune Response In Vitro ◽  
Selection Of

We have cultivated lymph node microfragments from ß-D-galactosidase (Escherichia coli) primed rabbits and have measured their secondary response directed towards the whole molecule (precipitating antibodies) and to a single determinant (activating antibodies) of the antigen. By decreasing the size of the fragments to 105 cells, we began to observe heterogeneity among identical cultures in terms of positivity of response, antibody specificity, and titers. The affinity of "early" activating antibodies was inversely proportional to the dose of challenge. While no maturation was seen in low and excessive challenge, in all cultures receiving intermediate doses the association constant was raised several orders of magnitude within periods of 20 days. The relevance of these data to the mechanism of affinity selection of antigen-sensitive cells is discussed.

scholarly journals INDUCTION AND REVERSAL OF IMMUNE PARALYSIS IN VITRO

1970 ◽  
Vol 132 (5) ◽  
pp. 845-857 ◽  
Author(s):  
V. S. Byers ◽  
E. E. Sercarz
Keyword(s):  
Immune Response ◽  
Cell Proliferation ◽  
Lymph Node ◽  
Cell Population ◽  
Antibody Production ◽  
Cellular Environment ◽  
Immune Paralysis ◽  
Antigen Removal ◽  
The Postponement

Induction of the immune response can only be completed after antigen is removed from the cellular environment. Primed rabbit lymph node fragments were cultured in vitro with 5 mg/ml BSA. If antigen was removed from the fragments 2 hr later, they produced a normal anti-BSA response, which was first evident 5 days later. If antigen removal was delayed for 3 days, the onset of the response was postponed for 2 to 3 days. Pulses with BUDR marked the periods of cell proliferation in both sets of cultures, and established that the postponement of antibody production was preceded by a postponement in the wave of proliferation among precursors of antibody forming cells. The similarity in avidity of antibody-containing fluids from normal and postponed cultures support the idea that the same cell population produced the response in each case. It was concluded that a reversible state of paralysis could be instituted in antigen-responsive cells, and this state did not depend upon cell-killing. The widespread incidence of temporary paralysis as an early aspect of the immune response was discussed.

scholarly journals ANTIGEN RECOGNITION: IN VITRO STUDIES ON THE SPECIFICITY OF THE CELLULAR IMMUNE RESPONSE

1969 ◽  
Vol 130 (5) ◽  
pp. 1031-1045 ◽  
Author(s):  
Stuart F. Schlossman ◽  
Judith Herman ◽  
Arieh Yaron
Keyword(s):  
Immune Response ◽  
Lymph Node ◽  
Cellular Immune Response ◽  
Lymph Node Cell ◽  
Antigen Receptors ◽  
Lymph Node Cells ◽  
Cellular Immune ◽  
Conventional Antibody ◽  
Sensitive Population

Studies of the immunochemical specificity of antigen-induced thymidine-2-14C incorporation in lymph node cells obtained from animals immunized to a series of closely related α-DNP-oligolysines, ϵ-DNP-oligolysines, and oligolysines have shown that the sensitized cell exhibits an extraordinary degree of specificity for antigen. The sensitized cell is maximally stimulated by the homologous immunizing antigen and can discriminate among compounds which differ from one another only in the position of a dinitrophenyl group or D-lysine residue on an identical oligolysine backbone. These studies support the view that the immunogen is not degraded prior to the induction of the immune response, and that the majority of cells produced as a consequence of immunization have stereospecific antigen receptors for the DNP-oligolysine used to induce the response; a smaller and more variably sized population of cells is produced with receptors specific for the oligolysine portion of the immunizing antigen. When specifically sensitized lymph node cell cultures are stimulated in vitro by heterologous DNP-oligolysines, the oligolysine- and not the DNP-oligolysine-sensitive population of cells appears to play a crucial role in the specificity of such cross-reactions. It is concluded from these studies that the antigen receptor on the sensitized lymph node cell differs in both kind and degree from conventional antibody. The chemical nature of the receptor and the means by which this receptor reacts with antigen to initiate the biosynthetic or proliferative cellular immune response still remain undefined.

scholarly journals Obesity impairs resistance to Leishmania major infection in C57BL/6 mice

2018 ◽  
Author(s):  
Franciele Carolina Silva ◽  
Vinicius Dantas Martins ◽  
Felipe Caixeta ◽  
Matheus Batista Carneiro ◽  
Graziele Ribeiro Goes ◽  
...  
Keyword(s):  
Immune Response ◽  
Lymph Node ◽  
Leishmania Major ◽  
Parasite Burden ◽  
Public Health Problem ◽  
Obese Mice ◽  
Obese People ◽  
Diet Induced Obesity ◽  
Draining Lymph Node

AbstractAn association between increased susceptibility to infectious diseases and obesity has been described as a result of impaired immunity in obese individuals. It is not clear whether a similar linkage can be drawn between obesity and parasitic diseases. To evaluate the effect of obesity in the immune response to cutaneous L. major infection, we studied the ability of C57BL/6 mice submitted to a high fat and sugar diet to control leishmaniasis. Mice with diet-induced obesity presented thicker lesions with higher parasite burden and more inflammatory infiltrate in the infected ear when infected with L. major. We observe no difference in IFN-γ or IL-4 production by draining lymph node cells between control and obese mice, but obese mice presented higher production of IgG1 and IL-17. A higher percentage of in vitro-infected peritoneal macrophages was found when these cells were obtained from obese mice when compared to lean mice. In vitro stimulation of macrophages with IL-17 decreased the capacity of cells from control mice to kill the parasite. Moreover, macrophages from obese mice presented higher arginase activity. Together our results indicate that diet-induced obesity impairs resistance to L. major in C57BL/6 mice without affecting the development of Th1 response.Author SummaryThe obesity is a public health problem and it is reaching extraordinary numbers in the world and others diseases are being involved and aggravated as consequence of obesity. What we know is that some diseases are more severe in obese people than in normal people. We did not know how obesity changes the profile of immune response to infectious agents, leading to the more severe diseases. That‘s why we decided to investigate how obese mice lead with Leishmania major infection. Leishmaniasis is a protozoa parasite infection considered a neglected disease. To try our hypothesis we gave a hipercaloric diet to induce obesity in C57BL/6 mice. After that, we injected L. major in the mice ear and followed the lesion for 8 weeks. We observed a ticker lesion and the cells from draining lymph node from obese mice produced more IL-17 than cells from normal mice. We also infected in vitro, macrophages from obese mice and stimulated the cells with IL-17, and we observed that the macrophages from obese mice are more infected by the L. major and it is worst in the presence of IL-17. Our results suggest that diet induced obesity decrease the resistance to infection.

Immune response profiles in vaccinated and non-vaccinated high- and low-responder mice during infection with the intestinal nematode Trichinella spiralis

Parasitology ◽  
1995 ◽  
Vol 110 (1) ◽  
pp. 71-78 ◽  
Author(s):  
K. Robinson ◽  
T. Bellaby ◽  
D. Wakelin
Keyword(s):  
Immune Response ◽  
Lymph Node ◽  
Mesenteric Lymph Node ◽  
Trichinella Spiralis ◽  
Serum Antibody ◽  
Specific Antigen ◽  
Mouse Strains ◽  
Mesenteric Lymph ◽  
Genetically Determined

NIH and C57 BL/10 (BIO) mice show genetically determined differences in their response to Trichinella spiralis infection. This study examines the influence of these on parameters of the immune response to infection after vaccination using muscle-larval excretory–secretory antigen in Freund's complete adjuvant. Serum antibody levels were greatly elevated when mice of both strains were vaccinated prior to infection; however, NIH produced significantly higher-level antibody responses than B10. Vaccination accelerated and increased the capacity of mesenteric lymph node T-cells to proliferate in vitro in response to specific antigen stimulation in both mouse strains but, in general, the stimulation indices of NIH cells were higher than those of the B10. The capacity of mesenteric lymph node cells (MLNC) and spleen cells (SC) to produce IL-5 and γIFN was measured after specific in vitro stimulation and early γIFN secretion was noted in the supernatants of NIH MLNC and SC, but not in B10 SC. Concentrations of IL-S rose steadily over the first 10–14 days after infection in cell cultures from both strains. Prior vaccination of these animals appeared to enhance cytokine levels. It is postulated that the efficacy of vaccination in NIH mice is a consequence of their genetically determined capacity to produce early and high-level responses to the antigens of T. spiralis and to express these in intestinal effector mechanisms.

scholarly journals Bacille Calmette-Guérin infection in the mouse. Regulation of macrophage plasminogen activator by T lymphocytes and specific antigen.

1978 ◽  
Vol 147 (4) ◽  
pp. 1175-1188 ◽  
Author(s):  
S Gordon ◽  
Z A Cohn
Keyword(s):  
T Lymphocytes ◽  
Plasminogen Activator ◽  
Macrophage Activation ◽  
Specific Antigen ◽  
Peritoneal Macrophages ◽  
Bacille Calmette Guerin ◽  
Vitro Assay ◽  
Purified Protein Derivative ◽  
Peritoneal Cells

High levels of plasminogen activator (PA) were induced in mouse peritoneal macrophages by infection with BCG, 2-6 X 10(7) viable organisms intravenously, followed 3-4 wk later by intraperitoneal challenge with purified protein derivative (PPD) 2 days before harvest. Macrophages obtained from infected animal without boosting showed little fibrinolytic activity, but challenge of Bacille-Calmette-Guèrin (BCG)-primed peritoneal cells with PPD in culture also enhanced macrophage PA 4- to 10-fold. Stimulation of macrophage PA by PPD depended on specifically sensitized thymus-derived (T) lymphocytes because it was abolished by pretreatment of BCG-primed peritoneal cells with anti-thy 1.2 antiserum and complement. A direct assay was developed in which nylon wool separated sensitized lymphocytes and PPD induced PA in macrophages from uninfected animals under defined conditions on 125I-fibrin. Enhanced macrophage fibrinolysis was proportional to concentration of PPD and the number of sensitized lymphocytes transferred. An indirect two-stage assay was also used to show that BCG-sensitized peritoneal cells released a soluble inducer of macrophage PA into the culture medium, after challenge with PPD. Induction of macrophage PA by PPD challenge in vitro made it possible to study the generation and activity of sensitized peritoneal lymphocytes at different stages of infection. Our results show that nonadherent peritoneal cells of BCG-infected mice provide a rich source of specifically sensitized lymphocytes and that macrophage activation is limited by continued availability of antigen, as well as sensitized lymphocytes. Induction of macrophage PA provides a sensitive, versatile, and rapid in vitro assay to study the role of lymphocytes and specific antigen in macrophage activation by BCG.

Mesenteric Lymph Node Tγδ Cells Induced by Gastrectomy in Mice Suppress Cell-Mediated Immune Response In Vitro via Released TGF-β

2007 ◽  
Vol 142 (1) ◽  
pp. 66-71 ◽  
Author(s):  
Andrzej Gryglewski ◽  
Pawel Majcher ◽  
Krzysztof Bryniarski ◽  
Stanislaw Konturek ◽  
Maria Ptak ◽  
...  
Keyword(s):  
Immune Response ◽  
Lymph Node ◽  
Mesenteric Lymph Node ◽  
Mesenteric Lymph ◽  
Cell Mediated Immune Response ◽  
Immune Response In Vitro

scholarly journals Macrophage-lymphocyte interaction. II. Antigen-mediated physical interactions between immune guinea pig lymph node lymphocytes and syngeneic macrophages.

1975 ◽  
Vol 141 (1) ◽  
pp. 138-154 ◽  
Author(s):  
P E Lipsky ◽  
A S Rosenthal
Keyword(s):  
Lymph Node ◽  
Guinea Pig ◽  
Dna Synthesis ◽  
Lymphocyte Proliferation ◽  
Specific Antigen ◽  
Physical Interaction ◽  
Reversible Binding ◽  
Immune Lymphocytes ◽  
Physical Interactions

The effect of specific antigen on the development of physical interactions between lymph node lymphocytes (LNL) obtained from animals which had been immunized to that antigen and macrophages was examined. We found that the presence of antigen, either limited to the macrophage () or free in the medium, profoundly increased the degree of ) or free in the medium, profoundly increased the degree of Mphi-LNL interaction observed. This enhanced interaction was dependent on the coincidence in the cultures of Mphi bearing antigen and LNL from animals specifically immunized to that antigen. Although antigen-independent interactions developed equally well between syngeneic and allogeneic combinations of lymphocytes and macrophages, antigen mediated interactions required that macrophages and lymphocytes be syngeneic. Prolongation of antigen-mediated Mphi-LNL interactions resulted in the induction of LNL DNA synthesis, initially involving those lymphocytes physically associated with antigen-bearing Mphi. These studies are interpreted to indicate that physical interaction between immune lymphocytes and antigen-bearing Mphi represents a morphological correlate of the functional activation of immune lymphocytes. Further, it is suggested that the physical events involved in lymphocyte proliferation may proceed sequentially from antigen-independent reversible binding of lymphocytes by macrophages to prolonged antigen-stabilized interaction eventuating in the triggering of specifically immune lymphocytes.

Dual phosphorylation in response regulator protein PrrA is crucial for intracellular survival of mycobacteria consequent upon transcriptional activation

2017 ◽  
Vol 474 (24) ◽  
pp. 4119-4136 ◽  
Author(s):  
Alok K. Mishra ◽  
Shivraj M. Yabaji ◽  
Rikesh K. Dubey ◽  
Ekta Dhamija ◽  
Kishore K. Srivastava
Keyword(s):  
Transcriptional Activation ◽  
Response Regulator ◽  
Regulatory Protein ◽  
Peritoneal Macrophages ◽  
Physical Interaction ◽  
Fast Growing ◽  
Regulatory Circuit ◽  
Regulator Protein ◽  
Promoter Dna

The remarkable ability of Mycobacterium tuberculosis (Mtb) to survive inside human macrophages is attributed to the presence of a complex sensory and regulatory network. PrrA is a DNA-binding regulatory protein, belonging to an essential two-component system (TCS), PrrA/B, which is required for early phase intracellular replication of Mtb. Despite its importance, the mechanism of PrrA/B-mediated signaling is not well understood. In the present study, we demonstrate that the binding of PrrA on the promoter DNA and its consequent activation is cumulatively controlled via dual phosphorylation of the protein. We have further characterized the role of terminal phospho-acceptor domain in the physical interaction of PrrA with its cognate kinase PrrB. The genetic deletion of prrA/B in Mycobacterium smegmatis was possible only in the presence of ectopic copies of the genes, suggesting the essentiality of this TCS in fast-growing mycobacterial strains as well. The overexpression of phospho-mimetic mutant (T6D) altered the growth of M. smegmatis in an in vitro culture and affected the replication of Mycobacterium bovis BCG in mouse peritoneal macrophages. Interestingly, the Thr6 site was found to be conserved in Mtb complex, whereas it was altered in some fast-growing mycobacterial strains, indicating that this unique phosphorylation might be predominant in employing the regulatory circuit in M. bovis BCG and presumably also in Mtb complex.

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