Hyperprolactinemia has been reported in some patients with active systemic lupus erythe matosus (SLE). To determine if there was an association between selected autoantibodies and hyperprolactinemia, we assayed prolactin concentrations in sera from women sub mitted to a reference antinuclear antibody laboratory. Autoantibody-positive samples were separated into groups that contained antibodies to double-stranded DNA (anti-DNA), anti bodies to SSA/Ro (anti-SSA/Ro), or antibodies to both SSA/Ro and SSB/La (anti-SSA/Ro- SSB/La). Results were compared with autoantibody-negative sera from age-matched women, submitted to the same laboratory. We also compared the study groups with a separate cohort of 84 healthy women who were not referred for autoantibody testing. Elevated prolactin levels were clustered in 20% of sera from anti-DNA-positive women ≤50 years of age. Twenty-one percent of anti-SSA/Ro-SSB/La-positive women < 50 years of age were hyperprolactinemic. Four of the 15 hyperprolactinemic women identified in this survey had no known cause of elevated prolactin. In the other 11 individuals, secondary causes such as hypothyroidism, pregnancy, chronic renal failure, and medications may have accounted for high serum prolactin values. We also examined sera by Western blot, to determine if immunoblot patterns were associated with elevated serum prolactin concentra tions. The hyperprolactinemic sera yielded novel bands migrating at 70 kd, 32 kd, and 16.5 kd. This study confirmed the reported associations of hyperprolactinemia with SLE and Sjögren's syndrome. Multiple factors appeared to contribute to elevated serum prolactin levels in women with connective tissue diseases, and the presence of hyperprolactinemia was related to unique findings on immunoblot analysis.

Abstract: Systemic lupus erythematosus (SLE), a chronic autoimmune illness, is influenced by hormones. High prolactin concentrations were associated with early death from autoimmune renal disease in NZB/NZW mice, an animal model of severe SLE. NZB/NZW mice that delivered and nursed pups and those that underwent pseudopregnancy had changes in serum IgG and autoantibodies. NZB/NZW mice treated with the prolactin‐suppressing drug bromocriptine had prolonged lives. Elevated serum prolactin concentrations are reported in SLE patients of both sexes. We found four women with long‐standing hyperprolactinemia who developed SLE. A survey of premenopausal women whose sera were submitted for autoantibody testing showed that 20% with anti‐ds‐DNA antibodies also had high prolactin levels. Many hyperprolactinemic patients whose sera were referred to an endocrinology laboratory had positive FANA tests (women 33%, men 53%) but did not have SLE. Disease activity was suppressed in six of seven SLE patients treated with bromocriptine. All had elevated disease activity and five became unexpectedly hyperprolactinemic after treatment stopped. Manipulating serum prolactin affords a means of treating clinical SLE activity.

OBJECTIVE To investigate the efficacy of bromocriptine in suppressing active systemic lupus erythematosus (SLE) in a therapeutic trial. METHODS We conducted an open label investigation of bromocriptine treatment in 7 patients with active non-life threatening SLE. Patients received bromocriptine daily during the treatment phase of 6 to 9 months and were followed for 5 months after bromocriptine was discontinued. Disease activity was assessed by determination of the SLE activity Measure (SLAM) and the Toronto SLE Disease Activity Index (SLEDAI). Serum prolactin concentrations and a battery of serologic and urine tests were obtained at baseline and at monthly intervals during and after bromocriptine treatment. RESULTS Serum prolactin concentration was suppressed from (mean +/- SEM) 11.2 ng/ml +/- 1.9 to 3.1 ng/ml +/- 1.7 after 6 months of bromocriptine treatment. The mean pretreatment SLAM score was 11.3 +/- 0.9;6 months of bromocriptine treatment significantly decreased the mean SLAM score to 6.0 +/- 1.6 (p = 0.03 compared to pretreatment measure). The mean SLEDAI score decreased from 16.0 +/- 2.0 to 5.9 +/- 0.8 (p = 0.02) during the same period. Bromocriptine treatment was associated with transient suppression of anti-dsDNA, and serum cholesterol was reduced significantly through the treatment period. After bromocriptine was discontinued, all patients had increased disease activity associated with rising serum prolactin concentrations. CONCLUSION These findings justify controlled trials to study the efficacy of bromocriptine in treating patients with active SLE.

MRL/lpr mice were treated with anti-CD4 mAb to define the role of CD4+ T cells in the pathogenesis of autoimmune disease and the lymphoproliferation characteristic of the strain. Anti-CD4 treatment was not associated with adverse effects, and survival of treated mice was increased over that of rat IgG-treated controls. Renal function was preserved, and the histologic severity of glomerulonephritis was minimal in treated mice. Lymphoid tissues of mice receiving anti-CD4 were effectively depleted of CD4+ T cells, and lymphoproliferation was markedly reduced. Serum IgG, anti-Sm, and anti-dsDNA levels were reduced significantly, while serum IgM and IgM rheumatoid factor levels were unaffected by anti-CD4 treatment. These data show that in MRL/lpr mice lymphoproliferation, renal disease, anti-Sm and anti-dsDNA antibody production, and elevated IgG levels are all linked to CD4+ T cell function. In contrast, both total IgM and IgM rheumatoid factor production appear to be the result of B-cell activity that is not regulated by CD4+ T cells.

Anti-histone antibodies have been reported in a number of human autoimmune diseases, most notably idiopathic and drug-induced lupus erythematosus. In the current study, anti-histone antibody activity was detected using ELISA and electroblotting techniques in sera from autoimmune NZBW, MRL-lpr, and MRL-+/+ mice. Anti-histone activity increased with age, maturing earlier in females, in both NZBW and MRL-lpr mice. Testosterone treatment decreased anti-histone activity in NZBW mice and estrogen treatment from 2 weeks of age increased anti-histone activity in MRL-lpr mice, suggesting that gonadal hormones modified the expression of autoantibodies recognizing these protein antigens. Estrogen also increased serum IgG levels in MRL-lpr mice. Sex hormones affected expression of antibodies recognizing soy milk proteins but not ovalbumin in a similar manner. Nitrocellulose Western blots of SDS gels probed with sera from both types of autoimmune mice most often demonstrated reactivity with histone1. Some mice, usually mature females, also recognized histone4, histone3, and histone2.

The immunosuppressive properties of cyclophosphamide prevent formation of anti-DNA antibodies and prolong lifespans in autoimmune NZB/NZW mice, an animal model of systemic lupus erythematosus. In the current study, NZB/NZW mice were treated with weekly doses of cyclophosphamide to determine if intermittent pulses of the drug were effective therapy. Life-long treatment with cyclophosphamide, 56 mg/kg/week, was started at the mean age of 6 weeks; results were compared with saline-injected control mice. Pulse therapy with cyclophosphamide suppressed anti-DNA antibody levels, prevented severe qlomerulonephritis and prolonged longevity. Seventeen of 19 treated mice developed neoplasms; 7 of these immunosuppressed animals had 2 to 4 separate neoplasms. Examination of earlier studies in this laboratory in which NZB/NZW mice were treated each day with cyclophosphamide showed that daily and weekly therapeutic regimens had similar immunosuppressive and oncogenic effects.

Autoimmune female New Zealand Black/New Zealand White mice were treated with frentizole, an experimental immunosuppressive drug. Three groups of "young," 8-week-old mice received high-dose frentizole (80-84 mg/kg/day), low-dose frentizole (8 mg/kg/day), or no drug (controls); these mice were followed until spontaneous death. Three groups of "old," 24-week-old mice with established lupus-like disease were treated with high or low doses of frentizole. Old control mice received no drug. After 12 weeks of therapy, surviving old mice were killed. Beneficial therapeutic response was achieved when high-dose treatment was started at an early age; antiDNA values were suppressed, and longevity was prolonged significantly. Frentizole did not arrest the progression of renal disease in old mice. Glomerulonephritis and vasculitis were the most common causes of death in young and old animals. Twenty-nine percent of young, high-dose-treated mice died with neoplasms. Large glomerular deposits of IgG, IgM, and C3 were present in renal tissue from treated and control mice. Peripheral lymphocyte counts and mitogenic responses of spleen cells were not changed by treatment. The efficacy of frentizole in a murine model of lupus supports its usefulness as an immunoregulatory drug.

An improved method of zinc administration, employing parenteral depot injection, was used as an experimental therapy in a mouse model of systemic lupus erythematosus. The depot method was used to produce a sustained increase in circulating plasma zinc levels for 7-day periods between injections. Female New Zealand Black/New Zealand White (NZB/NZW) mice were treated either with once weekly depot zinc, or with vehicle alone (controls) for 24 weeks. The zinc treatment produced a significant reduction in glomerular lesions. These data provide further evidence of the beneficial effect of zinc in inflammatory diseases.

Inbred Palmerston North (PN) mice are a newly recognized model of systemic lupus erythematosus. In this study PN mice with established autoimmune disease were treated until death with cyclophosphamide (8 mg/kg/day) or hydrocortisone (10 mg/kg/day). These doses had previously been found to prevent or suppress disease in another lupus model, the NZB/NZW mouse. In the PN strain, autoantibodies, severity of glomerulonephritis, and longevity were not influenced by treatment. Furthermore, the incidence of neoplasms was not increased in PN mice receiving prolonged therapy with immunosuppressive drugs. Unlike NZB/NZW mice, PN mice were resistant to the effects of Cyclophosphamide and hydrocortisone.

Frentizole is a benzimidazoleurea that has immunosuppressive properties in mice. Eleven steroid-treated patients with active systemic lupus erythematosus received frentizole (150-350 mg/day) in combination with stable or decreasing doses of prednisone in an open label trial. Nine patients completed at least one 21- to 75-day course of therapy with this drug. Clinical parameters of disease improved in 8 of these 9 patients. Mean DNA binding decreased by 28%, mean CH50 increased by 20%, and mean absolute lymphocyte and T cell counts decreased by 25-26%. Granulocytopenia was not observed. Three patients developed reversible hepatic toxicity. Clinical and serologic improvement was noted in 3 patients who accepted a second 90-day course of frentizole therapy.

Prolonged immunosuppressive therapy with cyclophosphamide increases the prevalence of neoplasms in NZB/NZW mice, an animal model of systemic lupus erythematosus. The current study was designed to compare the oncogenic properties of high dose cyclophosphamide with a low dose therapeutic regimen. Female NZB/NZW mice received life-long therapy with "high dose" cyclophosphamide, 16 mg/kg/day, or "low-dose" cyclophosphamide, 5.7 mg/kg/day; control mice received saline. High dose therapy clearly accelerated appearance of neoplasms. Seventeen of 19 mice treated with high-dose cyclophosphamide developed neoplasms at the mean age of 61 weeks. Fifty-seven percent of these tumors were mammary carcinomas. Neoplasms appeared in all mice treated with low dose; mean longevity in this treatment group was 80 weeks (compared to high dose treated mice, P less than 0.001). Carcinomas, pulmonary adenomas, and lymphomas were the most common tumors in mice receiving low dose therapy. Positive tests for ANA were suppressed in high dose treated mice. AntiDNA antibody levels and glomerulonephritis were decreased significantly in both groups of cyclophosphamide-treated mice compared to controls. It was concluded that the high daily dose of immunosuppressive drug was related to early oncogenesis in autoimmune NZB/NZW mice.

Deegan, Michael J., Walker, Sara Ellen, and Lovell, Sharon E.: Antibodies to double-stranded DNA. A comparison of the indirect immunofluorescent test using Crithidia luciliae and the DNA-binding assay. Am J Clin Pathol 69: 599–604, 1978. A new indirect immunofluorescent test for detecting antidouble-stranded DNA (dsDNA) antibodies was evaluated and the results compared with those of the DNA-binding test (modified Farr assay). The substrate, Crithidia luciliae (CL), is a nonpathogenic trypanosome containing a kinetoplast of dsDNA. The test was performed with fluorescein-conjugated polyvalent (anti-IgG, -IgA and -IgM) and monospecific anti-immunoglobulin antisera. Sera from 57 patients (75 specimens) with systemic lupus erythematosus (SLE), 30 patients with rheumatoid arthritis, 31 patients with scleroderma, 59 patients with other rheumatic diseases and 31 normal individuals were studied. Thirty-three of the 51 SLE sera with positive DNA-binding tests (binding > 20%) also had positive CL tests. The 18 CL-negative, DNA-binding-positive sera had DNA-binding values between 20 and 45%. AntidsDNA were usually IgG, often accompanied by IgM and occasionally IgA antibodies. Sera of eight patients who had conditions other than SLE were CL-positive, DNA-binding-negative. The CL test is a simple and reproducible method for detecting anti-dsDNA antibodies, but in its present form is not as sensitive as the DNA-binding test.

This report describes a previously unrecognized animal model of SLE, the PN mouse. Although outbred PN mice were studied originally as models of polyarteritis nodosa, their inbred descendants have autoimmune disease which closely resembles SLE. In the current study, positive indirect immunofluorescence tests for ANA appeared when the mice were 5 months old, and 80% of mice were ANA-positive at 10 months of age. Anti-DNA were detected in sera from newborn mice and from 53% of mice under 2 months of age. Seventy-six percent of PN mice developed anti-DNA at the age of 10 months. Glomerular deposits of IgG, IgM, IgA, and complement appeared at 2 to 4 weeks of age, and examination of renal tissue by electron microscopy showed basement membrane thickening and dense intramembranous deposits. Neoplasms arose in 14% of PN mice. Female mice died earlier than male mice, and the most common causes of death were glomerulonephritis and arteritis. It was concluded that the serologic and histologic characteristics of disease in PN mice resembled SLE.

This study investigated the effects of prolonged corticosteroid therapy on the course of spontaneous autoimmune disease and oncogenesis in NZB/NZW mice, an animal model of systemic lupus erythematosus. Twenty young female NZB/NZW mice were treated until death with low-dose hydrocortisone sodium succinate (3.3 mg/kg/day), and 21 mice received high-dose hydrocortisone (10 mg/kg/day). Fifteen control mice were injected with saline. Long-term therapy with either dose of hydrocortisone effectively prevented renal disease and prolonged lifespans in NZB/NZW mice. Fifty-six percent of low-dose treated animals developed neoplasms, and 38% of mice in this treatment group died with renal disease. Neoplasms caused death in 76% of mice receiving high-dose treatment. Long-term hydrocortisone therapy was associated with a predominance of sarcomas, which appeared in aged mice after a long period of treatment. In earlier studies conducted in this laboratory, cyclophosphamide treatment prolonged life in NZB/NZW mice. Ninety-seven percent of cyclophosphamide-treated mice developed neoplasms; most tumors were lymphomas or carcinomas. It was concluded that neoplasms occur commonly in old NZB/NZW mice with lives prolonged by immunosuppressive or antiinflammatory drugs. Nevertheless, the specific therapeutic agent used in each study influenced the types of neoplasms appearing in treated mice.