Andree A. Hoffman

Altered cellular immune response following peer separation

Abstract Lymphocyte response to phytohemagglutinin (PHA) and concanavalin A (Con A) mitogen stimulation was measured in a pair of pigtailed ( M. nemestrina ) monkey infants that had been raised together as peers since early infancy. at the age of 27 weeks, and following 3 baseline weekly blood samples, the infants were separated from each other for an 11 day period, and then reunited. A depression of lymphocyte stimulation by both PHA and Con A was noted during the latter part of separation and early reunion in both infants. The data support the notion that the disruption of an attachment bond produced by peer separation may impair these measurements of cellular immune function, and may be related to the increase in morbidity and mortality seen after bereavement.

The prognostic and therapeutic implications of DNA:Anti-DNA immune complexes in systemic lupus erythematosus (SLE)

Serum samples serially obtained from 50 patients with systemic lupus erythematosus (SLE) were studied for antibody to deoxyribonucleic acid (DNA) and circulating DNA:anti-DNA complexes during the active and inactive phases of their disease. The patients were divided into four categories: Group I: six patients without clinical evidence of central nervous system (CNS) or renal involvement. Group II: three patients with CNS lupus. Group III: nine patients with normal urinalyses and glomerular filtration rates, but morphologic evidence of glomerular disease. Group IV: 32 patients with overt lupus nephritis. Elevated anti-DNA levels were observed in 16 of 18 patients (88 per cent) in groups I, II and III during active disease. This persisted in 14 (77 per cent) during remission. DNA:anti-DNA complexes were demonstrated in four of 18 (22 per cent) during active disease and disappeared in all but one patient with progressive disease. In 30 of the 32 patients (94 per cent) in group IV, DNA binding was increased during active disease; this persisted in 21 (70 per cent) despite remission. Complexes were observed in 25 of the patients in group IV (78 per cent) with active disease. In six of these patients, complexes have persisted; two have died, one has progressed to renal failure and the remaining three patients continue to manifest active disease. This study suggests that measurement of DNA:anti-DNA complexes provides a valuable additional index of disease activity and prognosis in SLE.

The isolation and functional activity of polymorphonuclear leukocytes and lymphocytes separated from whole blood on a single Percoll density gradient

Abstract A simple, rapid, one-step density gradient technique was used which allowed the separation of functionally active lymphocytes and granulocytes from the same small quantity of whole blood. Whole blood was layered on a discontinuous gradient of Percoll and centrifuged. The granulocytes were collected at the 1.090 g/ml layer (70% Percoll) while the mononuclear cell (MNC) fraction was obtained at the 1.077 g/ml density (60% Percoll). Erythrocytes sedimented through the Percoll and were separated from the other cellular fractions. The cells at the 1.090 g/ml fraction contained 94% or greater granulocytes while the 1.077 g/ml fraction contained in general greater than 90% MNCs. While a wide range of percentage of recoveries was observed, an average of 72 and 63% was obtained for the granulocytes and MNCs, respectively. The recovered MNCs and granulocytes were assessed for their functional activity in several assay systems. The MNCs obtained from Percoll could be stimulated with mitogens to the same extent as MNCs obtained from a Ficoll-Hypaque gradient. Granulocyte functions as measured by chemotaxis, bactericidal activity, and superoxide anion generation were unimpaired after separation on Percoll. This method may be potentially useful in situations where one wishes to obtain multiple determinations of cellular immune function from small quantities of whole blood.

Cerebrospinal fluid and the choroid plexus during acute immune complex disease.

Abstract During acute immune complex disease in the rabbit, immune deposits could be detected in the choroid plexus, as well as the kidney, in a majority of the animals sacrificed from 9 to 14 days after the injection of bovine serum albumin (BSA). During this time there was a good correlation between deposits in the choroid plexus and increases in cerebrospinal fluid (CSF) albumin and IgG. A relatively unchanged CSF IgG to albumin ratio was observed, while the CSF to serum albumin level was increased, suggesting an alteration in the blood—CSF barrier permeability to serum proteins. In addition, the temporal appearance of BSA and anti-BSA in the CSF of these animals mirrored their appearance in the serum and in one animal immune complexes could be detected in the CSF. The significance of these results is discussed in relation to the pathogenesis of central nervous system (CNS) disease in systemic lupus erythematosus (SLE).

Antibodies to dissociated cerebellar cells in new zealand mice as demonstrated by immunofluorescence.

Abstract The presence of an antibody in New Zealand Black (NZB) mice to dissociated cerebellar cells from 6–10 day-old BDF1 mice was detected by indirect immunofluorescence. The sera from NZB mice displayed a significantly higher binding to cerebellar cells by this technique than did the sera from two non-autoimmune strains (BDF1 and CAF1). The NZB sera showed a positive correlation between an index of cytotoxicity to cerebellar cells and the percentage of cerebellar cells showing serum immunoglobulin (Ig) binding. The NZB sera also showed a significant positive correlation between IgM binding and cytotoxicity to cerebellar cells. No relationship was seen between IgG1 and IgG2 fluorescence and the index of cytotoxicity. In addition, in none of the mice was there a positive correlation between levels of thymocytotoxicity and binding by any of the tested classes of immunoglobulin to dissociated cerebellar cells. Further characterization of the antibody using Sephadex G-200 fractionation of NZB sera showed Ig binding to cerebellar cells for peaks I (void volume) and II of the three major peaks. Pooled normal mouse sera showed elevated Ig fluorescence in only the second peak. In all cases, the heightened fluorescence was determined to be IgM when the appropriate fluoresceinated antisera was used. The data suggest the presence, in some NZB mice, of an IgM antibody reactive with componets of the central nervous system.

Chronic Immune Complex Disease: Behavioral and Immunological Correlates

Using a fear avoidance paradigm, behavioral effects were seen in Sprague-Dawley rats in which chronic immune complex disease was induced. These effects were related to changes in urine protein that developed during the course of the experiment. Experimental animals also had glomerular deposits of rat gamma globulin and BSA as determined by immunofluorescence; C3 deposits were observed in half of these animals. BSA and/or rat γ-globulin, but not C3, was seen in the choroid plexus of half of the experimental animals. This is the first study to report behavioral changes associated with the induction of chronic immune complex disease in experimental animals.

Presentation of antigen by human newborn monocytes to maternal tetanus toxoid-specific T-cell blasts

The requirement for linked HLA-D antigen recognition for the proliferation of antigen-specific T-cell blasts provides a method for testing antigen presentation of human monocytes. We used maternal tetanus toxoid-specific T-cell blasts to show that human newborn monocytes can process and present antigen at least as well as maternal monocytes. These results suggest that human monocytes are relatively more mature at birth than those of newborn rats and mice. A major role for monocyte-macrophage immaturity in limiting the immune responses of human newborns is therefore unlikely.