It is well established that in susceptible mouse strains, chronic treatment with subtoxic doses of mercuric chloride (HgCl2) induces a systemic autoimmune disease, which is characterized by increased serum levels of IgG1 and IgE antibodies, by the production of anti-nucleolar antibodies and by the development of immune complex-mediated glomerulonephritis. Susceptibility to mercury is partly under the control of major histocompatibility complex genes. To study the susceptibility to mercury further, we investigated the in vivo effects of mercury in young autoimmune disease prone (NZB x NZW)F1 (H-2d/z) mice prior to establishment of spontaneous autoimmune disease. Mercury-susceptible SJL (H-2s) mice and mercury low-responder BALB/c (H-2d) mice were used as positive and negative controls, respectively. In (NZB x NZW)F1 mice, treatment with mercury stimulated an intense antibody formation characterized by increased numbers of splenic IgG1 and IgG3 antibody-producing cells as well as by elevated serum IgE levels. Injection with mercury also induced an increased production of IgG1, IgG2b and IgE antibodies in SJL, but not in BALB/c mice. The mercury-induced IgG1 response in (NZB x NZW)F1 and SJL mice was found to be polyclonal and autoantibodies against double-stranded (ds)DNA, IgG, collagen, cardiolipin, phosphatidylethanolamine as well as antibodies against the hapten trinitrophenol were produced. In addition, SJL, but not (NZB x NZW)F1 or BALB/c mice, produced IgG1 anti-nucleolar antibodies after treatment with mercury. Further studies demonstrated that (NZB x NZW)F1 and SJL mice developed high titers of renal mesangial immune complex deposits containing IgG1 antibodies 3 weeks after injection with mercury. Thus, a mouse strain genetically prone to develop spontaneous autoimmune diseases is highly susceptible to mercury-induced immunopathological alterations.

In this paper we have summarized our findings on immune activity in patients with Rheumatoid Arthritis. RA is characterized not only by the formation of various autoantibodies but also of a hyperreactivity of the B cell system, shown as an increased DNA synthetic rate of blood non-T, non-monocytic lymphocytes as well as an increased number of actively antibody secreting cells both in the blood and the synovial fluid. Synovial fluid contains biological activity which synergizes with PWM for the induction of Ig-secreting cells in blood from healthy controls. The factor can also substitute for T cells in the PWM-induced antibody synthesis in vitro. This activity fits well with the finding that SF contains a factor which induces increased formation of IgG in LPS-pretreated mouse cell cultures. Experiments show that the factor leads to a preferential increase in the production of IgG2b antibody secreting cells. Therefore, we conclude that synovial fluid contains a B cell differentiating factor with a selective effect on the induction of a particular IgG subclass.

The synovial fluid of patients with rheumatoid arthritis (RA) was found to contain IgG and/or IgG-containing immune complexes (ICs) that stimulated an intense antibody formation when injected into mice of certain strains, notably of NZ background. The response was characterized by high and sustained levels of IgG1 antibodies with rheumatoid factor (RF) activity. In the study described, we investigated whether it is the antibodies with RF activity in the synovial fluid, that are responsible for stimulation of mouse RF in vivo. Different mouse strains were injected with synovial fluid from a seropositive RA patient (RA-SF), with human monoclonal antibodies with RF activity, with a human non-RF monoclonal antibody or with different preformed RF-like antibody-antibody (Ab-Ab) ICs. The experimental mice were monitored subsequently for IgG1 RF production. IgG1 RF antibodies were found in all strains (NZB, BALB/c and CBA) injected with Ab-Ab ICs formed at equivalence, but only in NZB using RA-SF or human monoclonal antibodies with RF activity. Optimal production of IgG1 RF by Ab-Ab ICs required the integrity of Fc and F(ab)'2 portions respectively of the antibodies; soluble and truncated ICs were less effective. Further studies demonstrated that the IgG1 RF response was not simply the result of a specific immune response against human IgG, since humoral immunity against human IgG was induced only when combined with an efficient adjuvant. During a typical adjuvant-associated primary response specific antibodies of IgM, IgG1 and IgG2a isotypes were found, i.e. quite different from the selective IgG1 response induced by RF-like containing immune complexes. This conclusion is substantiated further by the clear differences in responses to IgG containing fraction obtained from RA-SF in NZ mice compared to other strains. Our findings argue for a different type of reaction leading to the selective IgG1 response and might aid in elucidating the mechanisms for chronic production of antibodies with RF activity in patients with RA.

Numbers of in vitro spontaneous IgG, IgM and IgA plaque-forming cells (PFC) as assessed by a modification of the protein A haemolytic plaque assay were determined in the blood and synovial fluid of patients with seropositive rheumatoid arthritis (RA) and compared with those of control groups. The total numbers of PFC were significantly higher in the peripheral blood of patients with active seropositive RA than in that of normal controls. In addition, most B lymphocytes in the synovial fluid of patients with active seropositive RA were active immunoglobulin (Ig) producers, whereas synovial fluid lymphocytes from patients with inactive seropositive RA and seronegative arthritis were not. In general, IgA PFC were relatively high in blood, whereas IgG PFC dominated in the synovial fluid. IgM PFC appear to be relatively low in blood and synovial fluid. However, a relative increase of IgG PFC was noted in the peripheral blood of patients with active RA. To test for polyclonality of the increased Ig synthesis, we tested the sera of patients and controls for the presence of polyclonal antibodies against sheep erythrocytes (SRBC) and SRBC modified by fluorescein isothiocyanate (FITC) and trinitrophenyl (TNP). No differences were observed with SRBC and TNP-SRBC agglutinin titres between patients and controls, but patients with RA had higher titres of FITC-SRBC agglutinins than normal sera. This finding supports the concept of a polyclonal nature of antibody production in RA patients.

3H-thymidine incorporation in cultures of peripheral blood mononuclear cells from patients with seropositive rheumatoid arthritis (RA) and healthy controls was measured in vitro in the absence of added stimulants. A significantly higher level of spontaneous DNA synthesis was found in cultures of mononuclear cells from patients with clinically active RA than from patients with inactive disease and normal controls. This activity was more apparent in 24-h cultures than in 72-h cultures. There was no correlation between DNA levels and IgM rheumatoid factor (RF) titres. When T- and non-T-cell populations were separated and cultured simultaneously with unfractionated cells, only non-T cells maintained high levels of DNA synthesis, and enrichment of surface membrane Ig+ (SmIg) cells was generally associated with enhancement of 3H-thymidine incorporation. Furthermore, no difference was found in spontaneous DNA synthesis between cultures either containing or depleted of phagocytic cells. Moreover, the addition of graded numbers of autologous monocytes to highly purified T- and non-T-cell populations did not alter the background DNA synthesis. Thus, endogenously activated cells in RA patients are neither T lymphocytes nor monocytes. A regulatory influence by monocytes could not be demonstrated. It is suggested that cells actively engaged in DNA synthesis in RA blood are non-T cells in origin, most probably B lymphocytes.