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Several types of IgG-dependent phagocytic stimuli independent of complement were investigated For their property to induct human polymorphonuclear neutrophil leucocyte (PMN) aggregation and adherence to human endothelial cells (EC) in culture, A Coulter counter method was employed for the detection of cell aggregation. Aggregated IgG. ovalbumin anti-ovalbumin (OV anti-OV) immune complexes (both insoluble and soluble) and opsonized latex particles induced a significant degree of PMN aggregation which was detectable as early as 2 mm alter exposure of PMN to these stimuli. This aggregation was dependent on divalent cations (Ca++ Mg++), The same phagocytic stimuli furthermore significantly increased adherence of PMN to cultured human EC and serum-coated plastic. Controls consisting of native IgG, and OV anti-OV complexes prepared From F(ab')2 antibody failed to induce either aggregation or increased adherence of PMN. These data suggest that exposure of PMN to IgG-dependent phagocytic stimuli induces increased adhesiveness of PMN and that interaction between the Fc-receptor of PMN and the Fe-portion of phagocytic stimuli is essential for this effect. [ABSTRACT FROM AUTHOR]

Intracytoplasmic inclusions containing immunoglobulin (Ig) and complement (C3) are found in normal neutrophils (PMN) after incubation with sera from patients with SLE. These inclusions are believed to be immune complexes removed by phagocytosis from the SLE patients' sera in vitro. Similar inclusions were also noted in the circulating PMN from some patients with SLE. In the present study we have examined the relationship between the presence of intracytoplasmic inclusions and various clinical and laboratory features of SLE. Blood from forty-five patients with SLE was drawn and separated at 37°C. Fresh heparinized blood was also obtained from normal volunteers and allowed to stand for 90 min at 37 C. The buffy coat cells from both normal and SLE groups were removed, centrifuged. washed and examined (direct method) or incubated in the SLE sera for 90 min at 37°C (indirect method). Slides of washed cells were prepared in the cytocentrifuge, stained with fluorescein-conjugated goat anti-human IgG, IgM, IgA and C3 and examined under ultra-violet light. By the direct method, 24% of patients had small intracytoplasmic inclusions in their neutrophils when stained for IgG suggesting that immune complexes were phagocytosed in vivo. None of twenty-one normal controls had similar inclusions. By the indirect method. 62% of SLE patients were positive for IgG, 15% for IgM, 8% for IgA and 31% for C3. None of the twelve normal controls were positive. By the indirect method, PMN inclusions containing both IgG and IgM correlated with clinical activity (P < 0.001). depressed serum complement (CH50, P = 0.026; and C3, P < 0.051). cryoglobulinaemia (P = 0.014). anti-nDNA antibodies (P < 0.001) and Clq-binding immune complexes (P = 0.008). A suggestive correlation with granulocytopenia was also observed. The presence of inclusions containing IgG alone did not correlate with any of these parameters. C3 and IgM appeared to be mutually exclusive, i.e. neither was present simultaneously. These findings suggest (1) that normal PMN on exposure to SLE sera develop intracytoplasmic inclusions by phagocytosis of immune complexes, (2) the presence of such complexes correlates with a number of parameters of disease activity, particularly when IgG and IgM are both present and (3) such complexes may be phagocytosed in vivo as suggested by the presence of inclusions in vivo and contribute to a number of granulocyte disturbances seen in patients with SLE. These abnormalities in granulocyte function may be important, predisposing factors for infection in patients with active SLE. [ABSTRACT FROM AUTHOR]

The relationship between renal interstitial mononuclear cell infiltration and renal tubule cell (TC) proliferation has been examined in (NZB × NZW)F1 hybrid (B/W) and control mice. TC proliferation was measured by tritiated thymidine ([³H]TdR) injection, autoradiographic examination of kidney sections and enumeration of labelled tubule cells. In B/W mice interstitial infiltration began at 5 months and reached a peak at 9 months. Proliferation of TC began at 7 months and also reached a peak at 9 months. Control mice consisting of NZB, CBA/J and C57B1/6J mice showed no increase with age. When the percentages of labelled TC in areas adjacent to interstitial infiltrates and distant from them were compared, the TC adjacent to infiltrates showed an approximately four-fold greater amount of labelling. Ultrathin light and electron microscopic sections demonstrated TC injury in areas in close proximity to infiltrates. These studies suggest that mononuclear cells in the interstitial infiltrate may he injurious to TC, leading to a sequence of injury and subsequent regeneration of these cells. In diseases in which interstitial nephritis is present, such as Sjögren's syndrome, the interstitial infiltrate may contribute to the observed TC dysfunction. [ABSTRACT FROM AUTHOR]

The effects of cyclophosphamide on antinuclear antibody levels, immune complex deposition in renal glomeruli, glomerular cell proliferation and glomerulosclerosis in the (NZB × NZW)F1 hybrid mouse have been investigated in order to better understand the mechanism of action of this drug on nephritis. Five groups of mice were injected daily with this agent (15 mg/kg) by the i.p. route over a 2-month period. Treatment periods began at 1, 3, 5, 7 and 9 months. Each group was killed at the end of the treatment period. During the last week [³H]Tdr (1 μCi/g) was injected daily i.p. Immune complex deposition was measured by quantification of glomerular immuno-fluorescent staining in both capillary loops and mesangium. Glomerular cell proliferation was quantified by measurement of the number of glomerular cells which incorporated [³H]Tdr during in vivo labelling. The number of [³H]Tdr-labelled cells in each of 100 glomeruli was counted using kidney autoradiographs of whole kidney slices. This technique provided a very reproducible and quantitative index of glomerular cell proliferation. Glomerulosclerosis was measured by determining percentage of PAS-positive material in each of 100 counted glomeruli. Immune complex deposition in the kidney preceded the glomerular cell proliferation. The proliferation reached a peak at 9 months of age. Glomerulosclerosis gradually increased with age until the animal's death. Cyclophosphamide significantly decreased antinuclear antibody levels, glomerular cell proliferation, and immunoglobulin staining in the glomeruli. Progression of glomerulosclerosis was significantly arrested. The present findings demonstrate a very significant effect of cyclophosphamide on three important pathological changes which occur in the nephritis of the (NZB × NZW)F1 hybrid mouse and provide information about possible mechanisms underlying these changes. [ABSTRACT FROM AUTHOR]

The possible rule of phagocytosis of circulating immune complexes by neutrophils in the production of the neutropenia of Felty's syndrome has been investigated. Normal neutrophils phitgocytosed massive inclusions from the sera from twelve of fifteen patients with Felty's syndrome when incubated with these sera. Such inclusions were phagocytosed from only three of fifteen patients with seropositive RA who did not have Felty's syndrome. Normal neutrophils were more effective than patient neutrophils with regard to phagocytosis of inclusions from the patients' serum suggesting a defect in phagocytic function of Felty's neutrophils. The titre of granulocyte-reactive antinuclear antibodies did not appear to be related to the degree of neutropenia. The data suggest that phagocytosis of circulating immune complexes by neutrophils may interfere with the function of these cells in combating infection and also render them susceptible to removal from the circulation thus leading to the development of neutropenia. [ABSTRACT FROM AUTHOR]

Prostaglandins, or related compounds, as well as estrogen and androgen are believed to be involved in the processes that lead to the development of murine lupus erythematosus (LE) in NZB/NZW F1 hybrid mice. In this investigation we measured NAD+-dependent 15-hydroxyprostaglandin dehydrogenase (PGDH) açtivity in kidney tissue of male and female NZB/NZW F1 hybrid mice and in kidney tissue of male and female mice of other strains. In each case the specific activity of PGDH was significantly greater in kidney tissue of male than in kidney tissue of female mice. However, the specific activities of PGDH in kidney tissues of males of various strains of mice were similar, and the specific activities of PGDH in kidney tissues of females of various strains-including the NZB/NZW F1 hybrid-were similar. Thus, while the lower activity of PGDH in kidney tissues of the female may be important in the pathogenesis of LE, the lower activity of this enzyme is not unique to the kidney of the female NZB/NZW F1 hybrid mouse. [ABSTRACT FROM AUTHOR]

The following differential effects of immunosuppressive therapy with Cyclophosphamide (CYCLOPH) and 6-mercaptopurine (6-MP) in the female NZB-NZW F1 hybrid strain have been observed: (1) CYCLOPH but not 6-MP significantly decreased antinuclear antibody level. (2) Both CYCLOPH and 6-MP significantly decreased glomerular cell proliferation. (3) Both CYCLOPH and 6-MP significantly arrested progression of glomerulosclerosis. (4) While CYCLOPH significantly diminished Ig deposition in the glomeruli, 6-MP had no effect on this phenomenon. (5) While CYCLOPH decreased subepidermal globulin deposition in the skin, 6-MP appeared actually to enhance subepidermal staining. Thus, the present studies demonstrated that CYCLOPH was superior to 6-MP in four of the five parameters studied. In the case of one parameter, Ig staining of the skin, 6-MP actually produced enhancement of the staining. Both CYCLOPH and azathioprine which is a derivative of 6-MP, are currently being used for the treatment of human SLE. The present findings suggest that of the two, CYCLOPH may be the drug of choice.

Human endothelial cells (EC) cultured from umbilical cord veins were reacted with sera from patients with systemic lupus erythematous (SLE), rheumatoid arthritis (RA) and normal persons. Immunofluorescent staining for IgG, IgM and IgA was then performed. Two types of control cells were also used, human foreskin fibroblasts and KB cells (a human carcinoma cell line). Sera from 9 of the 18 (50%) SLE patients showed cytoplasmic staining of EC for IgG. KB cells and fibroblasts did not stain. None of the RA or normal sera showed positive staining. In the 9 instances in which there was positive staining, 4 had evidence of cutaneous vasculitis. In contrast, none of the 9 patients without EC cytoplasmic staining had clinical evidence of cutaneous vasculitis. This data suggests that some SLE patients have an IgG antibody that reacts with EC cytoplasm which may be related to cutaneous vasculitis.

The pericardial effusion in a case of rheumatoid pericarditis was studied to determine whether immune complexes and mediators of cellular immunity, represented by migration inhibitory factor (MIF), were present. MIF-like activity was detected in the pericardial fluid, but only traces of immune complexes were revealed by ultracentrifugation. The MIF-like activity was partially characterized by column fractionation and sugar inhibition tests. The role of lymphokines in the pathogenesis of this case of rheumatoid pericarditis is strongly suggested.

Female B/W mice spontaneously develop an autoimmune disease that is similar to systemic lupus erythematosus. Antibodies to doublestranded DNA (dsDNA) and antinuclear antibodies develop in aging animals; death from immune complex-mediated glomerulonephritis occurs from 8 to 12 mo of age. It has been reported that prostaglandin (PG)E(1) treatment of such mice prolongs survival. In the present study, four groups of female B/W mice were studied beginning at 6-11 wk of age on the following regimens: (a) a synthetic diet that contained 20% safflower oil, (b) a standard laboratory chow diet, (c) a standard diet together with injections of PGE(1), and (d) an essential fatty acid-deficient synthetic diet that contained 20% coconut oil. All animals were tested monthly for antinuclear antibodies and anti-dsDNA. Kidney tissue was obtained for light and immunofluorescence microscopy when animals were dying. All disease manifestations were altered strikingly in the essential fatty acid (EFA)-deficient animals. Intermediate benefit was seen in PGE(1)-treated animals. 7% of the control animals and 18% of safflower oil-fed animals survived to 10 mo. In contrast, the PGE(1)-treated and EFA-deficient mice had a similar survival rate (78-88%). At age 16 mo, 78% of EFA-deficient mice and 45% of PGE(1)-treated mice were alive. 25% of the PGE(1)-treated and 55% of the EFA-deficient animals survived to 20 mo. Serum anti-dsDNA appeared at age 5 mo in safflower oil-fed and control animals, but not until 9 and 12 mo for PGE(1)-treated and EFA-deficient animals, respectively. All kidneys from 7- to 9-mo-old safflower oil-fed and control animals and the majority of kidneys from PGE(1)-treated animals were abnormal by light and immunofluorescence microscopy. Kidneys from EFA-deficient animals were essentially normal at 10 mo. At 13 mo, all PGE(1)-treated animals examined had significant kidney involvement, whereas none of the EFA-deficient animals had glomerulonephritis. These findings demonstrate that an EFA-deficient diet has a beneficial effect on murine lupus erythematosus.

23 of 42, or 55%, of patients with systemic lupus erythematous had immunoglobulin deposits along the epidermal basement membrane of uninvolved skin (positive lupus band test [LBT]). In patients with low serum complement levels, 91% had a positive LBT), as compared with 15% in those with normal complement levels. The LBT was positive in 70% of patients with clinical and laboratory evidence of renal disease, but in only 31% of patients without renal disease. 81% of patients with the more severe histologic forms of lupus nephritis, i.e., proliferative glomerulonephritis and membranous glomerulonephritis, and positive tests, whereas only 23% with mesangial glomerulitis or normal histologic findings were positive. Immunoglobulins of the same class found in the skin were detected in the glomeruli of patients examined by renal biopsy. These results suggest that there is a relationship between the occurrence of immunoglobulin in the epidermal basement membrane and the presence of the more severe forms of lupus nephritis.

Circulating polymorphonuclear leukocytes (PMN) from patients with psoriasis and psoriatic arthritis were found to be significantly more adherent to nylon fiber columns when compared with both normal and nonpsoriatic patient control groups. The increased adherence correlated positively with the extent of disease and was highest in those patients with psoriatic arthritis. Total leukocyte and PMN counts were increased in psoriasis patients and were highest in the psoriatic arthritis group. No increase in cell counts was found for mononuclear leukocytes. PMN adherence was not increased in lithium-treated patients or a nonpsoriatic patient control group although such patients did have significant granulocytosis. PMN's are frequently present in lesions of psoriasis as are activated complement components and abnormal keratinocyte cyclic nucleotide levels. These factors or others may cause a generalized activation of PMN's in psoriasis leading to migration of neutrophils into the skin lesion. The present study demonstrates a systemic effector cell alteration in psoriasis and contradicts the general concept that uncomplicated psoriasis is limited to the skin.

Sera from patients with untreated psoriasis were found to induce increased superoxide anion (O-2) generation when incubated with normal granulocytes (PMNs) and zymosan. Sera from patients receiving systemic chemotherapy induced O-2 generation which was similar to that of normal sera and significantly lower than sera from the untreated patients. O-2 production was measured by superoxide dismutase inhibitable ferricytochrome C reduction and was dependent on the presence of both zymosan and a heat labile serum factor. Serum C3c and C5 levels were elevated in both treated and untreated groups of psoriasis patients while C4 was elevated only in untreated patients. serum ceruloplasmin, a O-2 scavenger, was not decreased in patients with psoriasis, and consequently does not account for the increased O-2 generation. These data suggest that sera from patients with psoriasis have an increased capacity to activate PMNs. Activation of PMNs in cutaneous and joint lesions may play a pathogenic role in psoriasis.

The effect of cyclophosphamide on renal interstitial mononuclear cell infiltration and renal tubule cell (TC) proliferation has been examined in (NZB x NZW)F1 hybrid (B/W) and control mice. TC proliferation was measured by tritiated thymidine (3/Tdr) injection, autoradiographic examination of kidney sections, and enumeration of labeled tubular cells. Cyclophosphamide administered 2 months before sacrifice virtually obliterated interstitial infiltration and significantly decreased tubule cell proliferation at 3, 5, 7, 9, and 12 months of age.

Evidence has been found to document the presence of circulating immune complexes in all patients with Felty's syndrome. The sera of all 12 patients studied showed intermediate complexes by analytical ultracentrifugation. The sera of 9 of 12 patients (75%) showed precipitin lines upon immunodiffusion against IgM rheumatoid factor. Both findings were statistically increased above those in a matched group of patients with classic rheumatoid arthritis. The presence of circulating immune complexes in the sera of the Felty patients was consistent with the observation that large inclusions containing IgG, IgM, and complement were phagocytized by normal polymorphonuclear cells when incubated with sera of Felty patients. Normal polymorphonuclear cells phagocytosed inclusions from 77% of Felty sera, compared with 27% classic rheumatoid arthritis sera. It is suggested that the uptake of immune complexes by polymorphonuclear cells plays a role in the neutropenia of Felty's syndrome.

The effects of cyclophosphamide an antinuclear antibody levels, immune complex deposition in renal glomeruli, glomerular cell proliferation and glomerulosclerosis in the (NZB x NZW)F1 hybrid mouse have been investigated in order to better understand the mechanism of action of this drug on nephritis. Five groups of mice were injected daily with this agent (15 mg/kg) by the i.p. route over a 2-month period. Treatment periods began at 1, 3, 5, 7 and 9 months. Each group was killed at the end of the treatment period. During the last week [3H]Tdr(1 mu Ci/g) was injected daily i.p. Immune complex deposition was measured by quantification of glomerular immunofluorescent staining in both capillary loops and mesangium. Glomerular cell proliferation was quantified by measurement of the number of glomerular cells which incorporated [3H]Tdr during in vivo labelling. The number of [3H]Tdr-labelled cells in each of 100 glomeruli was counted using kidney autoradiographs of whole kidney slices. This technique provided a very reproducible and quantitative index of glomerular cell proliferation. Glomerulosclerosis was measured by determining percentage of PAS-positive material in each of 100 counted glomeruli. Immune complex deposition in the kidney preceded the glomerular cell proliferation. The proliferation reached a peak at 9 months of age. Glomerulosclerosis gradually increased with age until the animal's death. Cyclophosphamide significantly decreased antinuclear antibody levels, glomerular cell proliferation, and immunoglobulin staining in the glomeruli. Progression of glomerulosclerosis was significantly arrested. The present findings demonstrate a very significant effect of cyclophosphamide on three important pathological changes which occur in the nephritis of the (NZB x NZW)F1 hybrid mouse and provide information about possible mechanisms underlying these changes.

A variety of neurological complications may occur in the various connective tissue and "collagen-vascular" diseases. Most of these complications are due to vasculitis affecting various sites in the central or peripheral nervous system. While the evidence for definitive vasculitis in SLE is not strong, small vessel damage usually is present in anatomic sites which correlate well with clinical features. Although patients with rheumatoid arthritis also may have vasculitis, neurological complications are usually related to nerve compression by rheumatoid nodules or the arthritic process itself. Considerable controversy exists regarding the accuracy of various diagnostic tests. While corticosteroids are the mainstay of therapy for these conditions, there are no definitive studies proving their efficacy.

We studied the inheritance of HLA haplotypes in 10 families with more than one member affected with adult onset rheumatoid arthritis (RA). The frequency of DR4 was 81% among these patients. Nine families had DR4 bearing haplotypes and homozygosity for DR4 existed in 4 families. In 6 of these families DR4 positive haplotypes were shared among affected members. All but one of the affected sibs shared at least one haplotype with their index case. Ten percent of the unaffected relatives had rheumatoid factor (RF). HLA-DR4 or associated genes appeared to confer susceptibility for RF production and development of RA. However, these haplotypes were inherited also by many sibs who did not develop any manifestations of disease.

Absolute numbers of B (IgG-, IgM-, and IgA-staining) and T lymphocytes (sheep erythrocyte rosette-forming cells) were determined in patients with systemic lupus erythematosus and other connective tissue diseases in cyclophosphamide-treated and noncyclophosphamide-treated patients and in control subjects. In patients receiving cyclophosphamide, all three types of immunoglobulin-staining varieties of circulating B lymphocytes were significantly decreased. At the same time the circulating T lymphocytes were also significantly reduced. In patients with scleroderma treated with therapeutic doses of cyclophosphamide and studied sequentially, the reduction in B lymphocytes occurred first, with eventual depletion of both cell types. In 2 patients, an early rebound increase in T cells occurred followed by a marked reduction. These data indicate that the immunosuppressive effects of cyclophosphamide may be associated with a reduction in both cell types.