Sporadic idiotypic cross-reactivities between antibodies and T helper cells: one example of aleatory expression of T cell idiotypes.

Idiotypic cross-reactivities between T and B cell receptors have been taken in the past to suggest VH-gene expression by both types of lymphocytes. More recently, after the molecular characterization of TcR, those observations were reinterpreted to indicate idiotypic network regulation, operating to select cross-reactive idiotypes in both B and T cell compartments. In support of these views, it has been shown that T cell expression of idiotypes is controlled by IgH-linked genes and markedly altered in B cell-deprived mice, and that T cells "learn" idiotype expression from the B cell compartment in the first few weeks of life.

Immunocompetent autoreactive B lymphocytes are activated cycling cells in normal mice.

Frequencies of B cell clonal precursors producing antibodies that react with mouse thyroglobulin, mouse erythrocytes, beef hemoglobin, KLH, and sheep erythrocytes were determined by limiting dilution analyses among small, resting lymphocytes, and among large activated cells from normal adult mice. While frequencies of clones reacting with external antigens were equally distributed in large and small B cells, most, if not all, autoreactive B lymphocytes were found in the large cell fraction. Analysis of antithyroglobulin hybridomas isolated from normal mice revealed dissociation constants ranging from 10(-6) to 5-6 X 10(-7).

The basis for major histocompatibility complex (MHC) and immunoglobulin gene control of helper T cell idiotopes.

BALB/c helper T cells, prepared against 2,4,6-trinitrophenyl (TNP)-derivatized syngeneic spleen cells, fail to recognize other hapten modifications [(4-hydroxy-3-nitrophenyl)acetyl, fluorescein isothiocyanate] of syngeneic cells, as well as TNP-derivatized cells from major histocompatibility complex-congenic donors. T helper cell interactions with "presenting" cells and "target" B lymphocytes are inhibited by monoclonal antibodies directed either to the hapten or to I-A molecules on target cells. Helper activity is also inhibited by monoclonal anti-idiotypic antibodies directed to an idiotope expressed by the TNP-binding myeloma protein MOPC460, but not by soluble TNP-protein conjugates.

The role of immunoglobulin receptors in "cognate" T-B cell collaboration.

The functional effects of anti-Ig antibodies have been investigated, using an experimental system where B cell activation is brought about by direct and specific interactions with T helper (Th) cells without participation of surface Ig receptors on the responding B cell. We have used Th cell lines and clones directed to class II major histocompatibility complex antigens of the responding B cells, and titrated into cooperative cultures either purified rabbit anti-mouse mu, or monoclonal mouse anti-delta antibodies. Both types of antibodies greatly enhanced B lymphocyte responses to suboptimal concentrations of functionally efficient Th cells, while they had no effect in cultures containing optimal Th:B cell ratios.

Functional and biochemical evidence for the recognition of T cell receptors by monoclonal antibodies to an immunoglobulin idiotype.

Sharing of "idiotypes" by T and B cells with similar nominal specificities has been extensively reported in functional assays. The recent molecular characterization of T cell receptors has led to the suggestion that such idiotypic mimicries could result from "network" selection of available T cell repertoires. Alternatively, the validity of the conclusions taken from those functional assays could be questioned.

Natural effector T lymphocytes in normal mice.

The "natural" T-cell activity in normal unimmunized mice was studied. By double-parameter fluorescence-activated cell sorter analysis, it was found that 5-10% of all splenic Lyt-2+ and L3T4+ lymphocytes are large, of which more than half are in mitotic cycle. In contrast with small resting cells of the same phenotype, activated (large) T cells isolated from normal mice are functional effector cells: L3T4+ large cells induce normal B lymphocytes into proliferation and antibody secretion, while large Lyt-2+ cells efficiently suppress B-lymphocyte responses.

Different functional subsets of cultured murine T cells express characteristic levels of adenosine deaminase activity.

The level of adenosine deaminase (ADA) activity in mouse T-lymphocyte cultures was studied under different growth-supporting conditions and in mixed lymphocyte culture-derived long-term T-cell lines and clones. Early after the initiation of in vitro culture, the levels of ADA (2000 U/mg) were similar in bulk cultures either depleted or not depleted in Lyt-2+ T cells. Enrichment for cytolytic T lymphocytes (CTL) obtained by addition of exogenous interleukin 2 (IL-2), was accompanied by a net decrease of ADA activity (110 +/- 15 U/mg).

Interactions of small B lymphocytes with unprimed noncytolytic T cells: dissociation between "presentation" and growth induction.

The accessory cell requirements in lectin-dependent triggering and growth of unprimed Lyt-2-T lymphocytes were analyzed by quantitatively comparing the ability of small B cells and peritoneal macrophages to either induce reactivity to growth factors or support growth. Lightly or nonirradiated small B cells were 15 to 30-fold less efficient as compared to T cell-depleted peritoneal cell populations, in the support of the lectin-stimulated Lyt-2-T cell proliferation. In contrast, lightly irradiated small B lymphocytes were quantitatively as efficient as macrophages in mediating lectin-driven Lyt-2-T cell proliferation, if relevant supernatants were added into culture.

Functional analysis of pokeweed mitogen-dependent cell interactions in murine spleen cells. I. Lack of B-cell mitogenicity and low frequency of effector helper T cells.

The nature of lymphocyte responses on addition of pokeweed mitogen (PWM) to normal murine spleen cells was studied in low cell density cultures. PWM, over a wide range of concentrations, stimulated proliferation in a set of cells roughly 10-fold smaller than the lymphocyte populations responding to either concanavalin A or lipopolysaccharide. PWM also induced a relatively small number of B lymphocytes in these cultures to mature to Ig-secreting plaque-forming cells (PFC).

Activation and growth requirements for cytotoxic and noncytotoxic T lymphocytes.

The number and nature of the "signals" required for lymphocyte activation have been so repetitively and academically discussed over the last 15 years that both the readers and the authors appear exhausted by such exercises. Yet, what may be considered to be the essential question, the basis for self-nonself discrimination, remains to be clarified. Since it has been established that clonal expansion and maturation to effector functions are brought about by polyclonally ("immunologically nonspecific") active factors, it is obvious that the crucial "step" in this context is the initial interaction of antigen with specific receptors of immunocompetent lymphocytes.

A quantitative assay detecting small numbers of effector helper T cells, regardless of clonal specificity.

We have developed a new assay for quantitative detection of all helper T cells that can induce normal B lymphocytes to proliferation and Ig secretion. To establish the optimal assay conditions, we have used cloned T helper cells of defined specificities that had previously been shown to activate normal B lymphocytes expressing the specific antigen(s) on direct cellular interactions. As shown in this paper, 'irrelevant' B lymphocytes--that is, those that do not express either antigen or restriction elements recognized by the effector helper T cells--can also be induced in the presence of appropriate concentrations of pokeweed mitogen which are not mitogenic for the 'target' B lymphocytes.

Internal complementarities in the immune system: regulation of the expression of helper T-cell idiotypes.

More than half of BALB/c helper T lymphocytes specific for 2,4,6-trinitrophenyl (TNP)-modified syngeneic spleen cells are inhibited in their proliferative responses to antigen-presenting cells and in their cooperation with B lymphocytes by monoclonal anti-idiotypic antibodies directed to a TNP-binding BALB/c myeloma protein (MOPC 460). This inhibition is specific for anti-TNP-self helper cells of BALB/c origin and is controlled by IgCh-linked genes, as it is not observed with CB.20 helper cells of the same specificity.

T cell-dependent B cell activation.

T cell-dependent induction of small, resting B lymphocytes requires direct recognition of antigen and/or I-A/E molecules on the B cell surface by the inducing helper cell, and it does not require the participation of Ig receptors on the responding B cell. Triggering B cell receptors, therefore, are either the I-A/E molecules themselves, or other structures with complementarities on helper cell membranes that become available for productive interactions upon I-A/E recognition. It would appear that signal delivery by such triggering receptors can be regulated by a membrane complex of molecules, involving immunoglobulins, Class II MHC molecules and other classes of receptors, which in selective and distinct manners control the quantitative levels of expression and/or availability of the relevant structures.

B lymphocyte activation upon exclusive recognition of major histocompatibility antigens by T helper cells.

This study has investigated whether exclusive recognition of I-A or I-E molecules on the B cell surface by T helper cells is sufficient to activate resting B cells. Lines and clones of long-term-cultured T helper cells with specificity for I-A or I-E antigens have been derived from mixed lymphocyte cultures between spleen cells from major histocompatibility complex (MHC)-congenic mouse strains. These cells were tested for helper activity and proved competent to induce resting B lymphocytes expressing the specific MHC antigens to polyclonal expansion and maturation to Ig secretion.

Differential requirements for activation and growth of unprimed cytotoxic and helper T lymphocytes.

The requirements for activation and growth of T lymphocytes capable of mediating either cytolytic activity or help to B lymphocytes were studied in unprimed splenic T cell populations. The selectivity of expression of Lyt-2 antigens, the reactivity to soluble concanavalin A (Con A), to partially purified interleukin 2 (IL 2, T cell growth factor[s]) and to lectin-pulsed macrophages (M phi) were used in this analysis. Lectin-dependent cytotoxicity assays and a novel method that allows for the detection of all effector helper cells, regardless of their clonal specificities, were used for the functional identification of the responding T cells.

Activation of helper T cells for B lymphocytes in primary mixed lymphocyte cultures.

Normal B10.BR (H-2k, C57B1/6 background) spleen cells, enriched in primary mixed lymphocyte culture (MLC) for antigens of C3H/Tif mice (H-2k, C3H background), induced normal C3H/Tif but not B10.BR B lymphocytes to proliferate and produce Ig.

From the mechanisms of lymphocyte activation to internal activity in the immune system.

The principles of lymphocyte activation were summarized for all three sets of immunocompetent cells: B cells, cytotoxic cells and helper T lymphocytes. They were then used to derive the basic mechanisms and specificities which drive internal activity in the normal immune system and which select available antibody repertoires. It was postulated that "natural antibodies" are induced by "natural helper cells" and are selected on the basis of their idiotypic profiles, which are complementary to available T-helper-cell repertoires.

Distinct helper activities control growth or maturation of B lymphocytes.

A clone (C-11) of C3H/HeJ Lyt-1+2-T cells with specificity for "minor" antigens of C3H/Tif has been isolated which, in contrast to other similarly derived clones, did not activate polyclonal plaque-forming cell (PFC) responses in T cell-depleted "target" spleen cells. This clone, however, showed unaltered proliferative responses to the naturally occurring antigen(s) on presenting cells, and strongly synergized with regular helper clones in the induction of PFC responses. Further analysis demonstrated that C-11 cells are competent to stimulate extensive "target" B cell proliferation, but lack the ability to produce (or participate in the production of) maturation factors for activated B cells.

Differential macrophage requirements for T helper cell and T helper cell-induced B lymphocyte proliferation.

Major histocompatibility complex-restricted helper T cell clones against "minor" antigens expressed on B cell and macrophage surfaces, when confronted with appropriate T cell-depleted spleen cells, are induced to proliferation and, in turn, activate "target-responder" B cells to polyclonal growth and maturation. Irradiation of helper cell populations, however, demonstrates that their effector functions (and B lymphocyte responses) are independent of proliferative activity. Adherent cell depletion on Sephadex G10 columns, while completely abrogating helper T cell proliferation, does not abolish helper cell-induced B cell responses, demonstrating a remarkable quantitative difference in macrophage requirements for the growth of these two cell types.