Biomedical subjects
Publications and source records attributed to M W Hess.
This review attempts to summarize current knowledge of the cellular reactions observed during immune responses. A short presentation of organs and cells of the immune system is followed by a summary of the ontogeny of T and B cells and particularly of antigen receptors. Some examples of practical applications are quoted.
Primed mice with low titers of circulating tetanus antitoxin (AB) were stimulated via the hind footpads with either fluid tetanus toxoid alone (AG) to create in vivo complexes in AG excess, or the same dose of toxoid complexed at equivalence with isologous antibody (AB-AG CPX), to have in vivo complexes in AB excess. All experimental animals reacted with three topically distinct consecutive waves of enhanced proliferative activity in popliteal lymph nodes, i.e., in the T-zone (peak on day 2), in the medullary area, the main site of plasmocytopoiesis (day 3), and in lymphoid follicles (day 5-6). Maximum serum AB titers following injection of AG-AB CPX were only about 25% of those found in animals boosted with AG alone. This suppressive effect was best reflected in a comparable reduction in plasmocytopoiesis, and to an lesser extent in the proliferative activity within the T-zone, and not at all in the overall magnitude of germinal center formation and/or expansion. However, the patterns of germinal center kinetics differed markedly between the two groups: a high sharp peak of development on day 5, followed by a marked drop on day 6 characterized the response in mice given AG alone, and a broad peak around day 6 that of those receiving AG-AB CPX. These differences could not adequately be accounted for by variations in centroblast/centrocyte proliferation rate vs. pycnotic indices, so that different patterns of lymphoid cell emigration from the centers may be considered. The results suggest that immune complexes, fixed on follicular dendritic cells, with different antigen-to-antibody ratios have divergent effects on the development and kinetics of germinal centers, the principal sites of memory B cell generation.
Fluid free tetanus toxoid (FTT) alone or FTT complexed in vitro at equivalence (EQ) or in antibody excess (ABEX) with anti-toxin contained in a human gammaglobulin preparation (HGG), or HGG alone, were injected into the hind leg footpads of mice. Anti-toxin titres of mouse serum were measured and compared with proliferative reactions in popliteal lymph nodes, based on combined 3H-thymidine autoradiography and planimetry, as a function of time. FTT in complex with HGG in ABEX failed to elicit a measurable anti-toxin response but caused, of all the materials tested, the most marked numerical increase of germinal centres. This finding is in accord with results of earlier studies indicating that the same heterologous antigen-antibody complexes at EQ or in ABEX can prime the animals, usually without eliciting antibody production detectable by serum titration. The model system used in the present experiments is thus well suited for a separation of the two principal arms of the dichotomous humoral immune response, i.e. by inducing germinal centre and B cell memory development but not specific antibody formation.
The terminal involution pattern of the human thymus was studied based on autopsy cases (both sexes, age range 63-91 years). Large sections through the entire thymic fat body were examined with the help of both conventional histological and immunohistochemical techniques. The findings demonstrate that thymic atrophy in old humans (a) goes far beyond the degree of involution observed in small rodents; (b) results in a system of thin, branching, in part interrupted, non-keratinizing epithelial plates containing no typical Hassall bodies; (c) concerns all components of the thymus except fat tissue which progressively replaces original thymic structures; and (d) involves various types of disorganization of individual lobules with T and B lymphocytes often located outside rather than within epithelial remnants. Effects of low-level radiation on this final regression of the human thymus are unknown.
Murine spleen lymphocytes, cultured under serum-free conditions and stimulated with concanavalin A (Con A), showed an increased demand for supplemental concentrations of Ca2+ when 2-mercaptoethanol (2-ME) or L-cysteine were also present in the culture medium. The requirement for additional Ca2+ ions was particularly evident at high cell concentrations. The addition of SiO2 (Aerosil 200), in concentrations depending on the original level of Ca2+ in the culture medium, could substitute for the increased demand on calcium ions. This dependency of SiO2 effects on calcium levels in the medium was also evident when supraoptimal, i.e. inhibitory concentrations of silica were tested. Results of experiments with 45Ca suggest that silica particles may act as intercellular carriers of calcium. In contrast to spleen lymphocytes, Con A-stimulated thymus cells exhibited neither a need for an increase in the concentration of Ca2+ nor a dependency on the presence of 2-ME in the medium.
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This study was designed to answer the question if a primary immune response elicited in parathymic lymph nodes would influence the proliferative activity in the neighboring thymic cortex. Young adult mice were stimulated by an injection of aluminum phosphate-adsorbed tetanus toxoid into the peritoneal cavity which is known to be drained predominantly via parathymic lymph nodes. Animals were given an intravenous (i.v.) injection of deoxy(5-3H)cytidine and were killed 1 h later at various time points after priming. Combined morphometry, counts of cell numbers per unit area and radioautography revealed that a proliferative response of cortical thymocytes was confined to the vicinity of parathymic lymph nodes and peaked around day 14 after stimulation. It was also shown that the enhanced DNA synthetic activity took place in the subcapsular, outer zones rather than in the inner portions of the cortex. Results are discussed in the light of recent observations which demonstrated a translocation of small particles injected intraperitoneally (i.p.) into parathymic lymph nodes and the surrounding lymphatics, and from there, in small amounts, into the cortical parenchyma of the thymus. We conclude that cortical thymocytopoiesis can be enhanced in the segment adjacent to aprathymic lymph nodes as a result of i.p. injection of stimulants.
The present report deals with primary antibody responses to tetanus toxoid in 50-54-week-old ('ageing') as compared to 8-9-week-old ('young adult') mice. Antitoxin in the serum appeared 6 days earlier in the older than in the young animals, but in the latter reached 5 times higher titres on day 20. The magnitude of the proliferative response in the paracortex and the medulla of popliteal lymph nodes, as estimated by combined 3H-thymidine autoradiography and planimetry, was 3-7 times greater in the younger than in the older age group, thus approximately reflecting the difference in antibody titres on day 20. In contrast, germinal centre formation in response to the stimulus proved to be about 14 times less in ageing than in young adult mice. The findings demonstrate that, in the model system used, the age-related slopes of decline in humoral antibody responsiveness and proliferative reactivity in paracortex and medulla of first regional lymph nodes tend to be in parallel, while the ability of the immune apparatus to form germinal centres at this site deteriorates at a considerably faster pace. Results are also in line with the notion that centroblasts/centrocytes contribute little, if anything, to the ongoing antibody production elicited by the same stimulus which had triggered germinal centre formation. Finally, the observations made disprove the general validity of the suggestion that immune reactivity is maintained on the same level throughout life if tested with a novel antigen.
This is a short overview concerning possible relationships between immunodeficiency and cancerogenesis/leukemogenesis. Following introductory remarks on concomitant and sinecomitant antitumor immunity, various factors/mechanisms that could influence tumor-host-interactions are discussed, in particular properties of neoplastic cell lines, the microenvironment, cellular components of nonspecific resistance, and specific, i.e. antigen-directed, cell-mediated and humoral immune responses against cancer cells. The increased incidence of malignant neoplastic processes in patients with inherited or acquired immunodeficiency raises the question if a lack of antitumoral defense or ineffective antiviral immunity is more important. Available data indicate that once a cancer has reached a certain size, the chances for the host to reject it solely with the help of its immune apparatus are minimal. The possibility remains that immune reactions may be more efficacious against small numbers of immunogenic tumor cells, i.e. in the very earliest phase of a neoplastic process and when the cancer begins to metastasize.
The proliferation of mouse spleen cells and T-lymphocytes, initiated by the calcium ionophore A 23187 was studied by a serum-free culture technique. In contrast to Con A, A 23187 was capable of stimulating cells only if 2-mercaptoethanol, cysteine and glutathione (reduced form), respectively, were present in the culture medium. In the absence of one of these compounds a stimulating activity of A 23187 was observed only with high concentrations of cells (i.e., 10(7)/ml). With glutathione present, the cells could be stimulated only at concentrations of A 23187 which were found to be suboptimal in cultures with 2-mercaptoethanol. Human serum, fetal calf serum and bovine serum albumin shifted the active and optimally stimulating concentrations of A 23187 to higher values. A similar effect was observed with sera- and Con A-treated cells. The effect of sera and albumin was paralleled by a protecting effect of cells against high concentrations of A 23187.
The mechanisms regulating thymic lymphopoiesis are still a matter of debate. Intracortical proliferation and differentiation of thymocytes are thought to be controlled by locally produced humoral factors and close contact with epithelial, possibly also phagocytic, cells, and restricted by products of the major histocompatibility complex. The observation of a translocation of intraabdominally introduced PVP-coated silica particles (Percoll) via parathymic lymph vessels and through the thymic capsule into the cortical parenchyma demonstrates that the thymic cortex is accessible to materials carried with the transcapsular flux of interstitial fluid, and that this barrier is less effective than the blood-thymus barrier. The proliferative activity of cortical thymocytes following an intraabdominal injection of particulate tetanus toxoid was compared in sites adjacent to, and distant from, parathymic lymph nodes. Absolute numbers of DNA-synthesizing thymocytes were found to be much higher in cortical areas close to the lymph nodes, where lymphatic vessels are most numerous, than on the opposite sides of the thymic lobes. Taken together, these findings indicate that--in addition to intrinsic control mechanisms--cortical thymocyte production may be influenced by peripheral stimulation to some extent, and that materials from sites which are drained by parathymic lymph nodes may be important in this respect.
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DNA-synthetizing lymphocytes were identified in the lymph nodes regional and more distal to the site of developing P-815 tumours by incorporation of [3H]-thymidine followed by autoradiography of lymph node sections. It appeared that not only T but also B cell areas of draining and to a lesser extent of distal lymph nodes were stimulated by the growing tumour. This result was unexpected since neither humoral nor tumour cell-bound antibody could be identified so far as a functional correlate of B cell stimulation. In general the proliferative response of lymphocytes followed a biphasic pattern with an early peak of reactivity on days 2-3 and a second peak around day 12-15 after tumour cell inoculation. In the draining (axillary) lymph node the second peak of reactivity was suppressed, possibly as a consequence of metastatic tumour cells in this node when tumour cells were inoculated in the flank. The pattern of lymphocyte stimulation revealed larger individual variations after tumour cell inoculation in the flank than the foot pad. These results were associated with a slower and less regular drainage of carbon particles from the flank to the axillary and exceptionally the brachial lymph node than from the foot pad to the popliteal node after injection of India ink.
SiO2 particles enhanced the in vitro proliferation of splenic B cells obtained from BALB/c mice immunized with sheep erythrocytes. T cells from immunized animals were capable of replacing SiO2 with regard to the enhancement of both in vitro proliferation and the primary immune response. In addition, the stimulating effect of IgG, phytohemagglutinin (PHA) and concanavalin A (Con A) on spleen lymphocytes was also augmented by SiO2. The enhancing effect of SiO2 was higher at suboptimal than at supraoptimal concentrations of Con A. SiO2 is capable of binding Con A. The SiO2-bound lectin retained its mitogenic effect on spleen cells and, under certain conditions, SiO2-bound Con A was considerably more active in this respect than the free substance.
Regional labeling of mice by injection of cytidine-3H ([3H]CR) into the footpad of the left hind leg was used to evaluate lymphocyte traffic from the left regional nodes to the right popliteal lymph node (PLN) within a 24-h period, with or without concomitant primary or secondary stimulation of the right PLN with fluid tetanus toxoid. Results indicate that 1) in the case of primary antigen injection the relative contribution of lymphocytes from the left regional nodes to the small lymphocyte population present in the stimulated right PLN 24 h after labeling was slightly, but not significantly, greater than in non-stimulated controls; 2) a booster injection of antigen into the right hind leg footpad resulted in a significantly smaller relative contribution of lymphocytes from the previously primed left regional nodes to the small lymphocyte population in the right PLN, 24 h after injection of [3H]CR and secondary stimulation, as compared with controls or animals given a primary stimulation to the receiver node; and 3) in contrast to controls and mice subjected to primary stimulation only, the right PLN 24 h after booster contained a significant number of large lymphoid cells which, or whose precursors, had migrated to this site from contralateral nodes within a day, possibly also in the form of small lymphocytes. These findings are discussed in relation to the problem of lymphocyte recruitment and divergent behavior of non-committed lymphocytes as compared with memory cells in the initial phase after primary or secondary antigenic stimulation.
The translocation of Percoll microspheres (mean diameter 20-30 nm) from the intestinal lumen through the epithelial layer to internal organs was examined in suckling mice using transmission electron microscopy. Repeated administration of this material by gavaging for 7 consecutive days resulted in a heavy particle load of vacuolated enterocytes. A limited amount of Percoll was transported to the subepithelial tissue of both the villous mucosa and Peyer's patches where microspheres were found endocytosed, predominantly by macrophages. Even smaller numbers of particles reached mesenteric lymph nodes and, occasionally, milky spots of the omentum. Minor Percoll aggregates were easily found in Kupffer cells of the liver, indicating hematogenous translocation. Small numbers of particles were regularly detected in perivascular macrophages of the thymic cortex, which are in close contact with surrounding lymphocytes. We conclude that the thymic cortex is not totally inaccessible to particulate matter of the intestinal content.
We have studied cellular kinetic changes in the thymic cortex of ageing mice as a function of time after primary and secondary stimulation with aluminum phosphate adsorbed tetanus toxoid via the hind leg footpads. Absolute numbers of DNA synthesizing cortical thymocytes per cross-section of the thymus were computed from combined data obtained by planimetry, counts of cell density (cellularity) and assessment of the labelling index, 1 h after injection of (methyl-3H)-thymidine. The proliferative activity of these cells in the outer fourth of the thymic cortex reached a peak exceeding significantly control values by 45% 14 days after primary, but not secondary, peripheral antigenic stimulation. We conclude that the cortical thymocyte population responded in a distinct manner to the first immunization as opposed to the booster injection of the same antigen.