Deficiency in suppressor T cell activity in aged animals. Reconstitution of this activity by interleukin 2.

Spleen cells derived from aged (30 mo) C57Bl/6 mice are shown to be deficient in the ability to generate suppressor cell activity in vitro. The addition of IL-2 to cultures containing aged cells restores this function to a large extent.

Enhancement of T lymphocyte functions by Fc fragments of immunoglobulins. I. Augmentation of allogeneic mixed lymphocyte culture reactions requires I-A- or I-B-subregion differences between effector and stimulator cell populations.

Fc fragments derived from human Ig were found to be capable of enhancing T cell-mediated, antigen-induced proliferative and mixed lymphocyte culture responses. Maximum enhancement occurred when suboptimal amounts of antigen or suboptimal numbers of stimulator cells were employed. Augmentation of the allogeneic mixed lymphocyte culture reaction requires an I-A and/or I-B subregion difference between effector and stimulator cell populations. Although a significant proliferative response was observed with K- or D- region differences, Fc fragments were unable to enhance the response. The T cell population acted upon by Fc fragments in the potentiation of these responses bears the Lyt-1(+)23(-) phenotype.

Regulation of the immune response. I. The potentiation of in vivo and in vitro immune responses by Fc fragments.

Fc fragments derived from human and murine Ig were found to be potent adjuvants when administered with antigen. Both the in vivo and in vitro anti- sheep erythrocytes (SRBC) responses were significantly enhanced by Fc fragments. The adjuvant effect was shown to be extremely dependent upon the dose of antigen used, with the greatest enhancement occurring when suboptimal doses of antigen are employed. The anti-genicity of the Fc molecule was not related to its adjuvanticity because homologous Fc was as potent an adjuvant as heterologous Fc. Moreover, human Fc fragments enhanced anti-SRBC responses in mice which were tolerant to human gamma globulin.

Nuclear surface complex as observed with the high resolution scanning electron microscope. Visualization of the membrane surfaces of the neclear envelope and the nuclear cortex from Xenopus laevis oocytes.

The nuclear envelope and associated structures from Xenopus laevis oocytes (stage VI) have been examined with the high resolution scanning electron microscope (SEM). The features of the inner and outer surfaces of the nuclear surface complex were revealed by manual isolation , whereas the membranes facing the perinuclear space (the space between the inner and outer nuclear membranes) were observed by fracturing the nuclear envelope in this plane and splaying the corresponding regions apart. Pore complexes were observed on all four membrane surfaces of this double-membraned structure. The densely packed pore complexes (55/micron2) are often clustered into triplets with shared walls (outer diameter = 90 nm; inner diameter = 25 nm; wall thickness = aproximately 30 nm), and project aproximately 20 nm above each membrane except where they are flush with the innermost surface. The pore complex appears to be an aggregate of four 30-nm subunits. The nuclear cortex, a fibrous layer (300 nm thickness) associated with the inner surface of the nuclear envelope, has been revealed by rapid fixation. This cortical layer is interrupted by funnel-shaped intranuclear channels (120-640 nm diam) which narrow towards the pore complexes. Chains of particles, arranged in spirals, are inserted into these intranuclear channels. The fibers associated with the innermost face of the nuclear envelope can be extraced with 0.6 MKI to reveal the pore complexes. A model of the nuclear surface complex, compiled from the visualization of all the membrane faces and the nuclear cortex, demonstrates relations between the intranuclear channels (3.2/micron2) and the numerous pore complexes, and the possibility of their role in nucleocytoplasmic interactions.

Role of Fc receptors in lymphocyte activation: deficiency in T- and B-lymphocytes from aged animals.

Fc fragments derived from human immunoglobulin possess the ability to induce B-cell proliferation, polyclonal antibody responses, and augment cell-mediated and T-dependent humoral responses. However, aged animals display much lower responses to Fc fragment stimulation. Proliferation and polyclonal antibody synthesis are reduced two-five-fold in aged animals compared to the young-adult responses. Furthermore, Fc fragments are unable to potentiate plaque-forming cell (PFC) development in aged animals. Aged B-lymphocytes are deficient in responding to Fc fragments, as their admixture with young-adult T-cells does not restore polyclonal antibody formation. T-cells from aged animals are also ineffective in promoting the polyclonal response when mixed with young-adult B-cells. The T-cell lesion has been further defined as a deficiency in production of an Fc fragment induced T-cell-replacing factor [(Fc)TRF]. The inability of Fc fragments to potentiate anti-SRBC responses in aged animals is also due to a T-cell defect which can be corrected by supplementation with interleukin-2 (IL-2).

Characterization of host immunity during persistent vaginal colonization by Group B Streptococcus.

Streptococcus agalactiae (Group B Streptococcus, GBS) is a Gram-positive bacterium, which colonizes the vaginal tract in 10-30% of women. Colonization is transient in nature, and little is known about the host and bacterial factors controlling GBS persistence. Gaining insight into these factors is essential for developing therapeutics to limit maternal GBS carriage and prevent transmission to the susceptible newborn. In this work, we have used human cervical and vaginal epithelial cells, and our established mouse model of GBS vaginal colonization, to characterize key host factors that respond during GBS colonization. We identify a GBS strain that persists beyond a month in the murine vagina, whereas other strains are more readily cleared. Correspondingly, we have detected differential cytokine production in human cell lines after challenge with the persistent strain vs. other GBS strains. We also demonstrate that the persistent strain more readily invades cervical cells compared with vaginal cells, suggesting that GBS may potentially use the cervix as a reservoir to establish long-term colonization. Furthermore, we have identified interleukin-17 production in response to long-term colonization, which is associated with eventual clearance of GBS. We conclude that both GBS strain differences and concurrent host immune responses are crucial in modulating vaginal colonization.

The pattern of T lymphocyte differentiation is altered during thymic involution.

Thymic involution is likely to be a significant factor in the alteration of peripheral T lymphocyte function with age. The process of thymic involution involves the progressive loss of normal organ architecture and cellular composition, and a significant reduction in the output of mature T lymphocytes. The present study assesses the impact of thymic involution on the T cell differentiation process by quantitating the number and percent representation of various phenotypically distinguishable T cell developmental intermediates in C57BL/6 mice of various ages. The results suggest that several distinct sites in the developmental sequence are impacted by aging. By middle-age (14-17 months), significant perturbations in the frequencies of several CD4-CD8- (DN) subpopulations have occurred. These include a shift towards an increased percentage of Pgp-1+ IL-2R- DN cells, the earliest thymic progenitors, and a decreased percentage and total number of Pgp-1- IL-2R+ DN cells. Furthermore there is a threefold increase in the percentage of DN cells which express CD3 (from 16.6% to 45.5%) which occurs between 4 and 14 months of age. By 24-27 months of age, the percentage of the total DN population increases two- to threefold over that of young (2-3 months) animals, while the fraction of CD4+CD8+ (DP) is significantly reduced. These alterations are consistent with the possibility that thymic involution results in one or more 'developmental' blocks which limit key differentiative transitions within the DN population, and furthermore, the marked increase in the frequency of DN cells displaying CD3 argues that an alternative T cell differentiation pathway plays an increasingly significant role with advancing age.

Effects of the aged microenvironment on CD4+ T cell maturation.

Contributing to the functional alterations of the aged immune system is the accumulation of memory CD4+ T lymphocytes and decline in the proportion of naive cells occurring with advancing age. During attempts to alter the naive to memory ratio of CD4+ cells in aged mice, it was observed that regeneration of the peripheral T cell compartment resulted in a population which possessed the same memory cell-enriched characteristics as the unmanipulated age-matched controls. Thymopoiesis in aged mice does not appear to be altered in such a way as to give rise to emigrants with 'memory-like' characteristics. The aged peripheral microenvironment does, however, cause the accelerated maturation of mature, naive CD4+ T cells to the memory state.

Aging affects the regeneration of the CD8+ T cell compartment in bone marrow transplanted mice.

A chimeric mouse model has been used to determine the effect of aging on the differentiation of CD8+ T cells and on the regeneration capacity of the mature peripheral T cell pool after radiation induced depletion. Bone marrow cells from Thy 1.1+ mice were transplanted into lethally irradiated young or aged mice (Thy 1.2+). After 6 weeks, splenic CD8+ T cells were subjected to phenotypic and functional examinations by flow cytometry. Both young and aged mice were able to develop donor derived (Thy 1.1+) CD8+ T cells. Although the absolute number of T cells was reduced in aged recipients, the ratio of CD4+ to CD8+ T cells of donor-origin was the same in young Thy 1.1+ control mice as it was in both young and aged chimeric mice, indicating that aging has no effect on the ratio of CD4+ to CD8+ T cells produced by the thymus. However, the percentage of CD8+ cells in the total Thy 1.2+ (host-origin) T cell population was significantly higher in young chimeric mice than in age-matched Thy 1.2+ control mice (P < 0.01), suggesting that a significant over expansion of the Thy 1.2+ CD8+ subset occurred in young mice during regeneration. The Thy 1.1+ CD8+ T cells that developed in young hosts were of a naive phenotype with a majority of cells expressing a low level of CD44. In contrast, the majority of those that developed in the aged host displayed a memory phenotype with a high percentage of cells being CD44hi. In addition, the production of IL-4 and IFN-gamma by Thy 1.1+ CD8+ T cells was affected by the age of the host. A greater fraction of aged Thy 1.1+ CD8+ T cells could be induced to produce either IFN-gamma or IL-4 than young CD8+ T cells. These results suggested that the aged microenvironment has a significant effect on newly developed CD8+ T cells and that the age of the microenvironment also influences the regeneration capacity of CD8+ T cells.

T cell senescence.

The aging of the immune system, referred to as immunosenescence, is associated with a dramatic reduction in responsiveness as well as functional dysregulation. This deterioration of immune function with advancing age contributes to the increased incidence among the elderly of morbidity and mortality from infectious disease, and possibly autoimmunity and cancer. In mammals, the defense for fighting infectious agents is composed of the innate and adaptive immune systems. Macrophages, granulocytes, and natural killer cells are the major components of the innate system whereas T and B lymphocytes comprise the adaptive system. Although both compartments are affected, adaptive immunity is most susceptible to the deleterious effects of aging. Innate immunity functions immediately after birth and manifests little change throughout life. In contrast, adaptive immunity is immature at birth, peaks at puberty and progressively declines thereafter. Though marginal alterations in B lymphocytes are apparent, the dramatic decline in humoral and cell-mediated responses is predominantly the consequence of senescent T cells. The following review focuses on the aging effect on T cells as reflected in altered function, subset representation, development, lifespan and activation. Age-associated alterations in antigen presenting cells are also discussed since these cells are required for T cell activation and may impact T cell function.

Bioactive complement fragments in immunoregulation.

Several fragments derived from complement components have been identified as potent effector substances in in vitro assays that measure cell proliferation and antibody synthesis. The anaphylatoxin C3a suppresses the immune response but fails to influence T- or B-cell proliferation. The factor C5a augments both antibody production and antigen-induced, but not mitogen-induced, T-cell proliferation. C3a-mediated suppression occurs through the activation of a suppressor T-cell cascade with macrophage collaboration. C5a-mediated enhancement, depending upon the in vitro system studied, acts at the level of the helper T cell and/or macrophage. A fragment generated from treating iC3b with kallikrein (c3d-K) has aided in defining a structural region of the C3b molecule that can influence the level of circulating leukocytes. The factor C3d-K is also capable of suppressing both specific and non-specific T-cell proliferative responses and mitogen-induced B cell growth. The mechanism of C3d-K action is defined as a direct effect on "activated" T cells, even though IL-2 synthesis of treated cells is diminished. The effect of C3d-K is long lasting, non-reversible and requires only a short exposure to the target cell.

The age-dependent cytokine production by murine CD8+ T cells as determined by four-color flow cytometry analysis.

Flow cytometry has been used to simultaneously examine intracellular cytokine production and expression of CD44 and CD45RB by murine CD8+ T cells derived from young (2-3 months) or aged (18-22 months) mice. Cytokine production by aged CD8+ T cells differs from that by CD8+ T cells derived from young animals in that a significantly higher percentage of the aged can be triggered to produce interleukin (IL)-4, interferon (IFN)-gamma, and tumor necrosis factor alpha (TNF alpha). Conversely, a greater fraction of young CD8+ T cells produce IL-2. Aged mice not only have a higher percentage of CD8+/CD44hi T cells, but also a larger fraction of these cells are IFN-gamma+ and IL-4+, while a lower fraction are IL-2+ than is observed in young CD8+/CD44hi T cells. In terms of relative contribution to total cytokine synthesis, a greater fraction of CD8+ T cells produce IFN-gamma and IL-4 than in CD4+ T cells, whereas CD4+ T cells are the major producers of IL-2.

Toxicokinetics of Bordetella pertussis.

Pertussis vaccine immunization is required in most of the 50 states in the United States. Since it is required or recommended it is imperative that the clinician give this vaccine, crude as it is, in the safest possible environment. Giving specific active (verbal) as well as passive (written) instructions to the parent or caretakers is mandatory. If in doubt, defer or delete the pertussis portion.

Impaired responsiveness of IL-2 receptor-expressing T lymphocytes from aged mice.

IL-2 receptor-bearing splenic T lymphocytes derived from aged C57BL6/J mice (22-24 months) display a relative inability to respond to IL-2 when compared to similar cells from young (2-3 months) animals. As a population the aged cells incorporate less [3H]thymidine and fewer are able to undergo vigorous clonal growth. Both the CD4+ and CD8+ subsets display these defects. The clonal assay indicates that aged T cells, in addition to having longer cell cycle transit time, also have a higher frequency of cell cycle arrest than similarly activated young T cells. This defect in IL-2 responsiveness is distinct from those in early signal transduction which limit aged T lymphocyte entry into cycle and cannot be corrected by phorbol myristate acetate or ionomycin.

Early steps in T cell development are affected by aging.

Involution of the thymus accompanies aging, a process in which the organ diminishes in size and cellularity and becomes disorganized. The rate of T cell emigration from the thymus is markedly reduced with age, and phenotypic analyses have identified alterations in the relative proportions of the major thymocyte subpopulations. The present studies made use of the capacity of the thymus to regenerate following irradiation from an intrathymic radio-resistant precursor population. By analysis of the differentiation of this "wave" of thymocytes, it was determined that aging most severely affects the earliest developmental transitions. While the overall rate of differentiation does not appear to be affected in older mice, fewer thymic progenitors initiate differentiation. The reduced expansion of late pre-T cells in the middle-aged is due to the smaller pool size of these cells.

A factor that can be used to regulate an in vitro primary immune response.

A factor has been isolated that supports the stimulation of an immune response against heterologous erythrocyte antigens in mouse spleen cells cultured in a medium that contains deficient serum. The factor is secreted by certain spleen cells that are obtained in permanent culture after leukemia virus infection. The factor permits antigen-sensitive cells derived from bone marrow to mature to antibody-forming cells. This factor can be used in controlling the initiation of a primary immune response in vitro.

Cell-mediated immunity in aged mice: an underlying lesion in IL 2 synthesis.

It is well established that immune potential declines with age and that aged animals are unable to mount vigorous cell-mediated immune responses. Production of the lymphokine, Interleukin 2 (IL 2) in response to the T cell mitogen concanavalin A or allogeneic stimulation is also diminished in aged individuals. This lesion in IL 2 synthesis in aged mice underlies their deficiency in cell-mediated responses. Mixed lymphocyte responses, cell-mediated lympholysis, and antigen-induced T cell proliferation are all markedly enhanced by supplementation with this lymphokine.