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Reactive oxygen species (ROS) play important roles in cellular senescence and organismic aging. Furthermore, they have been implicated in some of the adverse effects of chronic stress due to elevated peripheral levels of catecholamines. Here, we applied three different techniques to individually compare the systemic and intracellular oxidative stress in aged (23 months) and young (5 months) Sprague-Dawley rats, and in young rats treated for 12 or 24 h with norepinephrine (NE). Thiol groups of blood serum proteins (RSH) were determined by means of Ellman's reaction. Intracellular ROS were assessed in spleen cells and peripheral blood lymphocytes (PBL) by carbonylation of cellular (spleen) proteins as determined by immunoblotting (Oxyblot) and/or by means of 2',7'-dichlorofluorescein (DCF) fluorescence. As compared to the young, untreated controls, both old rats and NE treated young rats showed similarly lowered RSH values paralleled by elevated intracellular ROS levels or enhanced Oxyblot signals. Individual RSH values were highly significantly, negatively correlated with respective Oxyblot data as well as with DCF fluorescence. The results confirm the roles of ROS in aging and adrenergic stress in the rat model, and suggest that the decrease in RSH of blood serum may be taken as a valid indicator for the enhanced oxidative stress in lymphocytes.

An apparent gradual decrease of IgG1 serum levels of up to 40% occurs within 48 h of storage at room temperature. The effect does not concern any other IgG subclass, and is more pronounced in sera of smokers. A linear correlation was found between the extent of this "storage effect" and the initial concentration of IgG1, which rules out an enzymatic process following Michaelis-Menten kinetics. PAGE and Western blots of density gradient separated serum proteins revealed the presence of noncovalent self aggregates of IgG1 in stored sera. Addition of superoxide dismutase prevented both the formation of aggregates as well as the decay of IgG1 values. It is concluded that the instability of IgG1 is due to an enhanced propensity of this molecule to form self-aggregates, whereby O2(-)-radicals play a functional role. This mechanism, however, is not relevant to a previously detected selective decrease of relative IgG1 levels in sera of patients afflicted with malignant diseases of various tissue origin.

The present study provides first evidence for the presence of a thymic cell growth factor (TCGF) in supernatants (20 hrs) of mitogen (Con A) stimulated chicken spleen cells. 2 out of 3 batches of supernatant prepared according to a procedure originally described for the mouse system (10) induced a vigorous proliferative response of Con A prestimulated chicken spleen cells without significant mitogenic effect on unstimulated lymphocytes. No crossreactivity of this chicken-TCGF was observed with prestimulated murine lymphocytes, nor did potent mouse-TCGF preparations exhibit any proliferative effect on chicken cells. The implications of these data for both phylogenetic as well as practical experimental aspects are discussed.

In contrast to systemic autoimmunity, spontaneous autoimmune thyroiditis of Obese strain (OS) chickens is associated with a marked T cell hyperreactivity in vitro, i.e. an increased proliferation and interleukin 2 (IL 2) secretion in response to Concanavalin A (ConA). In the present study we report an enhanced capacity of OS peripheral lymphoid cells (splenocytes and peripheral blood lymphocytes, PBL) to adsorb fluorescein isothiocyante (FITC) labelled ConA, but not phytohemagglutinin (PHA). However, the elevated ConA binding cannot be a prerequisite for in vitro ConA hyperreactivity as OS thymocytes are normal with respect to ConA binding but nonetheless exhibit elevated responses to this mitogen. Moreover, ConA binding does not correlate with the frequency of cells able to express IL 2 receptors upon short term ConA stimulation. The percentage of ConA activatable cells was found to be increased in OS- PBL as compared to normal control PBL, but was unaltered in OS splenocytes. This finding points to a further mechanism of T cell hyperreactivity in OS chicks in addition to the previously reported defects in nonspecific immunosuppression. Finally, enumeration of cells in the S phase revealed that enhanced proliferation of OS T lymphocytes was not restricted to the in vitro response to ConA and phytohemagglutinin (PHA) but also occurs in vivo.

Monoclonal antibody INN-CH 16 recognizes a surface determinant exclusively present on activated chicken T lymphocytes. The present experiments suggest that this chicken activated T lymphocyte antigen (CATLA) represents the chicken IL-2 receptor or an associated structure, as (i) the kinetics of CATLA expression during T cell activation are analogous to those described for the mammalian receptor for IL-2, (ii) the IL-2 dependent proliferation of mitogen prestimulated chicken lymphocytes is competitively inhibited by INN-CH 16, and (iii) pretreatment of T lymphoblasts with INN-CH 16 drastically reduces their capacity to absorb IL-2 activity from supernatants of mitogen activated chicken lymphocytes.