Antitrinitrophenyl (TNP) Plaque Assay. Primary Response of Balb/c Mice to Soluble and Particulate Immunogen

246

134

Antigens have been classified previously into three categories, thymus-dependent (TD), thymus-independent type (TI) 1, and TI-2, based upon thymic dependence and ability to stimulate an immunodeficient strain of mouse, CBA/N. Here we demonstrate that the different antigen classes elicit IgG antibodies of different subclasses. TD antigens stimulate predominantly IgG1 antibodies, with smaller amounts of IgG2 and IgG3 being expressed. TI-1 antigens stimulate almost no IgG1 antibodies and equal amounts of IgG2 and IgG3. TI-2 antigens elicit predominantly IgG3 antibodies. Mice expressing the CBA/N phenotype are known to be nonresponsive to TI-2 antigens. This was confirmed in this study. In addition, we demonstrate that the IgG3 component of the response to TI-1 antigens is virtually absent in mice expressing the CBA/N phenotype, which supports our previous finding that the CBA/N defect may be restricted to a B-lymphocyte subpopulation containing most of the precursors of IgG3-secreting cells.

Section: Antigen Preparation and Immunizations Trinitrophenyl-n-(amimentioning

confidence: 99%

Subclass restriction of murine antibodies. II. The IgG plaque-forming cell response to thymus-independent type 1 and type 2 antigens in normal mice and mice expressing an X-linked immunodeficiency.

Slack¹,

Der-Balian²,

Nahm³

et al. 1980

246

134

“…A modification of the hemolytic plaque technique was used to detect antibody secreting cells (12,13), using TNP-conjugated sheep erythrocytes as indicator cells (14). Secondary IgG anti-TNP plaque-forming cell (PFC) responses were assayed 7 days after cell transfer.…”

Section: Demonstration Of Pc-specific Helper T Cells In Nbf1 and Balbmentioning

confidence: 99%

Phosphorylcholine-specific helper T cells in mice with an X-linked defect of antibody production to the same hapten.

Kaplan¹,

Quintáns²

1979

F1 male mice with the CBA/N X-linked defect that are unable to produce plaque-forming cell responses to phosphorylcholine (PC) provide normal PC-specific helper T-cell activity when compared to F1 female littermates. Inhibition of helper activity with anti-idiotypic antiserum indicates that PC-specific T cells from both NBF1 female and male mice possess predominantly BALB/c myeloma protein HOPC-8 idiotypic determinants. Therefore, the CBA/N defect cannot be explained as a deletion of genes coding for V-region anti-PC specificities. The demonstration of helper activity in NBF1 male mice, which occurs in the absence of anti-PC antibody synthesis, also demonstrates the endogenous origin of the T-cell receptor.

“…Sheep erythrocytes (SRBC) were reacted with 2,4,6-trinitrobenzene sulfonic acid (Sigma Chemical Co., St. Louis, Mo.) according to the method of Rittenberg and Pratt (11). 50 ~l of an 8% suspension of TNP-conjugated SRBC were added to 0.5 ml of 0.5% agarose.…”

Section: Assay Of Pfc Animals Were Sacrificed By Cervical Dislocatiomentioning

confidence: 99%

Immunological studies of aging. II. Loss of IgG and high avidity plaque-forming cells and increased suppressor cell activity in aging mice.

Goidl¹,

Innes²,

Weksler³

1976

210

The magnitude and heterogeneity of the immune response to dinitrophenylated bovine gamma globulin was measured in aged and young mice at a cellular level using an inhibition of plaque-forming cell assay. The primary and secondary responses of 24-mo-old mice were markedly depressed in magnitude and restricted in avidity for the DNP determinant when compared to 2-mo-old animals. Bacterial lipopolysaccharide given at the time of immunization increased the restriction in heterogeneity seen in 12- and 24-mo-old mice. Indirect PFCs were more severely depressed than direct PFCs in 24-mo-old mice. Syngeneic, lethally irradiated, 2-mo-old mice reconstituted with aged spleen cells exhibit the depressed and restricted response to DNP-BGG seen in old mice. When 10(8) young thymus cells were given together with old spleen cells the heterogeneity of the response was increased. When 2-mo- and 24-mo-old spleen cells were transferred together into young recipients the magnitude of the response to the young spleen cells markedly reduced. Thus, there appears to be a loss of thymic-helper cells and an increase in suppressor activity in aged animals.