scholarly journals Mechanism of adjuvant activity of dental plaque: in vitro activation of residual helper T-cell precursors in T-cell-deficient murine spleen cell cultures.

1977 ◽  
Vol 17 (3) ◽  
pp. 567-571 ◽  
Author(s):  
P Chen ◽  
J J Farrar ◽  
J J Oppenheim ◽  
S E Mergenhagen
Keyword(s):  
T Cell ◽  
Spleen Cell ◽  
Cell Cultures ◽  
Dental Plaque ◽  
Helper T Cell ◽  
Adjuvant Activity ◽  
Murine Spleen ◽  
In Vitro Activation

scholarly journals H-2-restricted cytotoxic effectors generated in vitro by the addition of trinitrophenyl-conjugated soluble proteins

1978 ◽  
Vol 147 (2) ◽  
pp. 352-368 ◽  
Author(s):  
A Schmitt-Verhulst ◽  
CB Pettinelli ◽  
PA Henkart ◽  
JK Lunney ◽  
GM Shearer
Keyword(s):  
T Cell ◽  
Spleen Cell ◽  
Cell Cultures ◽  
Sodium Dodecyl ◽  
Soluble Proteins ◽  
Immunofluorescent Staining ◽  
Cytotoxic T Cell ◽  
Target Cells ◽  
Spleen Cells

Murine spleen cells from normal donors were cultured in vitro with trinitrobenzene sulfonate (TNBS)-conjugated soluble proteins, i.e., bovine gamma globulin (TNP-BGG) or bovine serum albumin (TNP-BSA). Addition of 100 μg of any of these TNP-proteins to the spleen cell cultures led to the generation of cytotoxic T-cell effectors which were H-2-restricted and TNP- specific. The lytic potential of such effectors was comparable to that generated by sensitization with TNBS-modified syngeneic cells, and was restricted to haplotypes shared at the K or K plus I-A, or the D regions of the H-2 complex. Greater effecter cell activity was generated by addition of TNP-BGG against TNBS-modified targets which shared K plus I-A than against modified targets which shared the D region with the responding cells, which suggests that the same immune response genes are involved when the response is generated by the addition of TNP-conjugated soluble proteins or of TNBS- modified cells. H-2-restricted, TNP-specific effecter cells were generated by culturing mouse spleen cells with syngeneic cells which had been preincubated with TNP- BGG or TNP-BSA for 1.5 h. The addition of unconjugated soluble proteins to the cultures did not result in cytotoxic effectors detectable on H-2-matched targets, whether the targets were prepared by modification with TNBS, or by incubation with either the unconjugated or TNP-conjugated proteins. Depletion of phagocytic cells in the tumor preparation by Sephadex G-10 column fractionation before incubation with TNP-BSA had no effect on their lysis by the relevant effector cells. Immunofluorescent staining of tumor target cells with anti-TNP antibodies indicated that TNP could be detected on the tumor cells within 10 rain of incubation with TNP-BSA. The cytotoxic response generated by addition of the TNP-proteins to spleen cell cultures was found to be T-cell dependent at the effector phase, as shown by the sensitivity of the lytic phase to absorbed RAMB and complement. Furthermore, the response did not appear to be attributable to antibody-dependent cellular cytotoxicity. Three mechanisms were considered which could account for the generation of H-2-restricted, TNP-specific, cytotoxic T-cell effectors by the addition of soluble TNP-proteins. These include covalent linkage of activated TNP groups from the soluble proteins to cell surface components, macrophage processing of the soluble conjugates and presentation to the responding lymphocytes in association with H-2-coded self structures, or hydrophobic interaction of the TNP-proteins to cell surfaces. Results obtained from sodium dodecyl sulfate gel patterns indicating that cell-bound TNP was still linked to BSA, and the observation that phagocytic-depleted cells could interact with the soluble TNP-proteins and function as H-2-restricted targets, appear not to favor the first two proposed mechanisms.

scholarly journals THYMUS-INDEPENDENT (B) CELL PROLIFERATION IN SPLEEN CELL CULTURES OF MOUSE RADIATION CHIMERAS STIMULATED BY PHYTOHEMAGGLUTININ OR ALLOGENEIC CELLS

1972 ◽  
Vol 136 (4) ◽  
pp. 962-967 ◽  
Author(s):  
P.-F. Piguet ◽  
P. Vassalli
Keyword(s):  
T Cell ◽  
Spleen Cell ◽  
Cell Cultures ◽  
Bone Marrow Cells ◽  
Spleen Cells ◽  
Radiation Chimeras ◽  
Lymphocyte Cultures ◽  
Mixed Lymphocyte Cultures

Spleen cell cultures of radiation chimeras (thymectomized, lethally irradiated mice repopulated with bone marrow cells and thymocytes bearing different chromosomal markers) were stimulated by phytohemagglutinin (PHA) and F1 allogeneic spleen cells. Karyotypic analyses showed a marked predominance of T mitoses on the 2nd and 3rd days of culture followed by a strong predominance of B mitoses on the 4th and 5th days. Analysis of cells undergoing their first mitoses showed that the majority of T mitoses on day 3 resulted from continuous T cell division, and that most cells entering their first mitoses at that time were of B type. Mixed lymphocyte cultures (MLC) of chimeras immunized against allogeneic spleen cells showed sometimes, but not always, a response different from "primary" MLC, with an earlier and stronger predominance of BM mitoses. The role of stimulated T cells in the induction of B mitoses was shown by (a) the incapacity of T-depleted spleen cells to be stimulated by PHA or in primary or secondary MLC, and (b) the restoration of the mitotic response of B cells to PHA by adding to the T cell-depleted culture either a very small number of T cell (identified by their different karyotype: "in vitro chimeras") or the cell-free supernatant of a 24 hr MLC.

scholarly journals IMMUNIZATION OF DISSOCIATED SPLEEN CELL CULTURES FROM NORMAL MICE

1967 ◽  
Vol 126 (3) ◽  
pp. 423-442 ◽  
Author(s):  
Robert I. Mishell ◽  
Richard W. Dutton
Keyword(s):  
Spleen Cell ◽  
Cell Cultures ◽  
Culture System ◽  
Cultured Cells ◽  
Inhibitory Effect ◽  
Initial Exposure ◽  
In Vitro Immunization ◽  
In Vitro Response

A culture system for cell suspensions from mouse spleens has been described. The system provides adequate conditions for in vitro immunization on initial exposure to heterologous erythrocytes. The in vitro response closely parallels that observed in vivo with respect to size, early kinetics, antigen dose, and the inhibitory effect of passive antibody. The response of cultured cells differs in two respects from that seen in vivo. There is an increase in the ability to discriminate between different varieties of homologous erythrocytes and the in vitro response does not appear to be limited by whatever mechanisms regulate the in vivo response.

IVIg prevents the in vitro activation of T cells by neutralizing the T cell activators

2013 ◽  
Vol 150 (1-2) ◽  
pp. 54-60 ◽  
Author(s):  
Lauriane Padet ◽  
Renée Bazin
Keyword(s):  
T Cells ◽  
T Cell ◽  
In Vitro Activation

scholarly journals T Cell-Mediated Delay of Spontaneous Mammary Tumor Onset: Increased Efficacy with In Vivo versus In Vitro Activation

2005 ◽  
Vol 174 (8) ◽  
pp. 4662-4669 ◽  
Author(s):  
Leigh A. O’Mara ◽  
Lyse A. Norian ◽  
Darren Kreamalmeyer ◽  
J. Michael White ◽  
Paul M. Allen
Keyword(s):  
T Cell ◽  
Mammary Tumor ◽  
Spontaneous Mammary Tumor ◽  
Tumor Onset ◽  
In Vitro Activation

scholarly journals JIA patient T cells differentiate into Th1, Th17 and Th1.17 effector cells under Th1 polarizing conditions

2021 ◽  
Author(s):  
Anna E Patrick ◽  
Tashawna Esmond ◽  
Kayla Shoaff ◽  
David M Patrick ◽  
David K Flaherty ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cell ◽  
Cell Cultures ◽  
Th17 Cells ◽  
Mononuclear Cells ◽  
Healthy Children ◽  
Th1 Cell ◽  
Peripheral Blood Mononuclear ◽  
Ifn Gamma

Objective. T helper cells develop into discrete Th1, Th2 or Th17 lineages that selectively express IFN-gamma, IL-4/IL-5/IL-13, or IL-17, respectively and actively silence signature cytokines expressed by opposing lineages. Our objective was to compare Th1, Th2 and Th17 polarization in cell culture models using JIA patient samples. Methods. Peripheral blood mononuclear cells were isolated from JIA or healthy prepubescent children. T cell naive and memory phenotypes were assessed by flow cytometry. T cell proliferation was measured using a fluorescence-based assay. Th cell cultures were generated in vitro and IFN-gamma, IL-17, and TNF-alpha measured by ELISA and flow cytometry. Results. JIA Th1 cells produced increased IFN-gamma and inappropriately produced IL-17. JIA Th17 cells produced increased IL-17. JIA Th1 cell cultures develop dual producers of IFN-gamma and IL-17, which are Th1.17 cells. JIA Th1 cultures expressed elevated levels of both T-bet and ROR-gamma-T. RNA sequencing confirmed activation of immune responses and inappropriate activation of IL-17 signaling pathways in Th1 cultures. A subset of JIA patient samples was disproportionally responsible for the enhanced IFN-gamma and IL-17 phenotype and Th1.17 phenotype. Conclusions. This study reveals that JIA patient uncommitted T cell precursors, but not healthy children, inappropriately develop into inflammatory effector Th1.17 and Th17 cells under Th1 polarizing conditions.

Cytotoxicity Is Not Required for the Anti-Leukemia Efficacy of Human H-Y-Specific CD4+ T Lymphocytes In Vivo.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 3553-3553
Author(s):  
Attilio Bondanza ◽  
Lothar Hambach ◽  
Zohara Aghai ◽  
Monica Casucci ◽  
Bart Nijmeijer ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Lymphocytes ◽  
Cell Clone ◽  
Scid Mice ◽  
Helper T Cell ◽  
T Cell Clone ◽  
Control Clone

Abstract Abstract 3553 Poster Board III-490 Introduction Minor histocompatibility antigens (mHag) play a major role in the graft-versus-leukemia (GvL) effect following HLA-matched allogeneic hemopoietic cell transplantation (allo-HCT). Clinically, the GvL effect coincides with the emergence of mHag-specific CD8+ cytotoxic T lymphocytes (CTL). Experimentally, targeting a single mHag with human CD8+ CTL has a major anti-leukemia effect in NOD/scid mice. Altogether, these observations suggest that mHag-specific cytotoxicity by CD8+ T cells is an important component of the GvL effect. In contrast, little is known on the contribution of mHag-specific CD4+ T cells. Female-to-male allo-HCT is characterized by a low rate of leukemia relapse, indicating that H-Y-encoded mHag are potent leukemia-regression antigens. Earlier, we described a DRB3*0301-restricted H-Y mHag epitope inducing CD4+ helper T-cell responses in H-Y-mismatched HLA-matched allo-HCT. Aim: The aim of this study is to elucidate the role of mHag-specific human CD4+ T lymphocytes on the GvL effect. Methods The ALL-CM leukemia cell line, derived from a male (i.e. H-Y+) HLA-A0201+, DRB30301+ patient, reproducibly engrafts in NOD/scid mice after administration of 10×106 cells. Both an HLA-A0201-restricted H-Y-specific CD8+ CTL clone and the DRB30301-restricted H-Y-specific CD4+ helper T-cell clone that we earlier described were used to investigate the anti-leukemia efficacy of CD8+ and CD4+ T cells in NOD/scid mice. Results In vitro, the CD8+ H-Y specific CTL clone was highly cytotoxic against the ALL-CM leukemia. The H-Y specific CD4+ helper T-cell clone did not lyse the leukemia, but produced IFN-γ upon recognition. Infusion of the H-Y-specific CD8+ CTL clone (25×106 cells/mouse) 3 days after ALL-CM leukemia challenge significantly delayed leukemia progression by 3 weeks compared to a CMV-specific CD8+ CTL control clone (p<0,001). Despite no measurable in vitro cytotoxicity, the H-Y-specific CD4+ helper T-cell clone (25×106 cells/mouse) delayed leukemia progression by 2 weeks compared to a leukemia non-reactive HLA-DR1-specific CD4+ helper T-cell control clone (p<0,001). In vitro co-incubation of the H-Y-specific CD4+ helper T-cell clone did not influence leukemia proliferation but induced up-regulation of MHC-class I and II, CD80, CD86 and CD40. In vitro, pre-incubation of leukemia cells with the H-Y-specific CD4+ helper T-cell clone irradiated did not improve the in vivo anti-leukemia efficacy of the H-Y-specific CD8+ CTL clone. Co-infusion of the H-Y specific CD4+ helper T-cell clone did not augment the in vivo persistence of the H-Y-specific CD8+ CTL T-cell clone. Nevertheless, the co-infusion resulted in a delay in leukemia progression of approximately 5 weeks, suggesting an additive, non overlapping anti-leukemia mechanism. Conclusions Minor Hag-specific human CD4+ T lymphocytes may contribute to the GvL effect through a direct, non cytotoxic mechanism, which could be additive to that of CD8+ CTL. The nature of this non cytotoxic GvL effect is currently under investigation. A.B. and L.H. equally contributed to this study. Disclosures: No relevant conflicts of interest to declare.

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