Heterozygosity at Gm loci associated with humoral immunity to osteosarcoma.

Serum samples from 50 Caucasian patients with osteosarcoma were tested for the presence of antibodies to osteosarcoma-associated antigens (OSAA) and typed for nine Gm markers. A highly significant association was found between Gm 3;5,13,14 and unresponsiveness to OSAA, and between 1,3,17;5,13,14,21 and responsiveness to OSAA. These results suggest the existence of complementary immune response genes which in the heterozygous condition permit a response to OSAA.

Altered heterogeneity of monocytes in acute myelomonocytic leukemia.

Human peripheral blood monocytes isolated from normal donors and patients with acute myelomonocytic leukemia (AMML) were separated on a discontinuous density gradient of bovine serum albumin (BSA) into five fractions. Cells from each fraction were assayed for cell surface markers, prostaglandin E2 (PGE2) production, ability to affect proliferation in response to antigen by autologous peripheral blood lymphocytes previously depleted of monocytes, and ability to regulate immunoglobulin (Ig) synthesis by allogeneic B-lymphocytes. Fractions 1-5 from normal donors contained 11, 10, 23, 34, and 22%, respectively, of the total number of monocytes. In contrast, in 6 patients with AMML fraction 3 was considerably larger (52%) than any other fraction, in 1 patient comprising 87% of her monocytes. Cells from each fraction differed markedly in accessory function. In general, cells from fraction 3 were poorer as helper cells than cells from other fractions. They also produced after stimulation larger amounts of PGE2 than did cells from other fractions of the gradient. These data show that PBL contain a subpopulation of monocytes, which either helps poorly or suppresses in vitro immunologic function of T-cells (proliferation) and B-cells (lg synthesis), and that this subpopulation is increased in the blood of patients with AMML.

Activated T cells in type I Charcot-Marie-Tooth disease: evidence for immunologic heterogeneity.

Common recognized variability in the familial peripheral neuropathy, type I Charcot-Marie-Tooth disease (CMT I), led to an examination of cell-mediated immune responses in 23 CMT I patients. Increased numbers of activated T cells were found in the peripheral blood of 14 (61%) patients using fluorescent monoclonal Ta1 antibody as quantitated by flow cytometry. Altered immunoregulation was also suggested by increased levels of prostaglandin-mediated lymphocyte suppression. In the other nine CMT I patients, immune responses were normal. Lack of a relationship between Ta1 expression and CMT clinical symptoms, but with consistency within six CMT families, support the concept of immunologic heterogeneity in type I CMT with a possible genetic component.

Expression of Schwann cell and peripheral T-cell activation epitopes in hereditary motor and sensory neuropathy.

To evaluate possible immune-mediated mechanisms in hereditary motor and sensory neuropathy (HMSN-I, Charcot-Marie-Tooth syndrome), we examined class II major histocompatibility complex antigen expression (MHC-II, HLA-DR) in Schwann cells and peripheral lymphocyte T-cell (Ta1, CD26) activation in five unrelated adults with HMSN-I. Evidence of increased activation expression was found in both compartments but the pattern did not suggest a general state of hyperimmunity or appear related to clinical characteristics of HMSN. Significantly increased CD26+ T-cell activation and greater than normal fluctuation of values occurred intermittently in sequential tests of eight HMSN patients and at single time points in 24 others. The combined data, consistent with repeated stimulations of an immune reaction under normal feedback control, suggest that HMSN-I expresses some characteristics also found in autoimmune polyneuropathies.

Activated retinitis pigmentosa peripheral lymphocytes adhere to and alter cultured human retinal pigment epithelial cells.

Interleukin-2 receptor (IL-2R) is an activation molecule that, when expressed on peripheral blood lymphocyte (PBL) membranes, indicates the secretion of IL-2 and initiation of an immune system activation cascade. Comparing the average of IL-2R expression in 34 patients with retinitis pigmentosa (RP) syndrome (561 +/- 282 cells/mm3; mean +/- standard deviation) with 35 age-matched normal subjects (194 +/- 39 cells/mm3), it was found that those with RP had greater numbers of IL-2R-positive cells (P less than 0.001). The increased amounts of IL-2R on PBL of 29 RP and the homotypic self-aggregation of RP PBL by phase and scanning electron microscopy led to the study of the interaction of RP PBL with cultured human postmortem retinal pigment epithelial cells (RPE). A direct correlation was found between the amount of IL-2R expression and the numbers of RP lymphocytes adhering to RPE monolayers. However, the adherence effect was not unique to RP syndrome but appeared to be a nonspecific result of lymphocyte activation. Greater adherence to RPE than normal also was observed in PBL from disease control subjects with elevated IL-2R values and in PBL stimulated by the mitogen, concanavalin A (Con-A). In addition, RPE monolayers were destroyed by Con-A-stimulated PBL that showed 95-98% IL-2R expression. Similar, but less serious effects, occurring in RPE cells after 1 wk's cocultivation with RP PBL, suggested that activated RP lymphocytes can be cytotoxic to RPE during prolonged contact. Because macrophage-like cells and class II major histocompatibility complex expression have been found in RP-affected retinas, immune-mediated cytopathologic effects may contribute to retinal degeneration in RP.

Acquired spontaneous cytotoxicity of K562 in a subpopulation of monocytes.

Human peripheral blood monocytes were placed on a discontinuous density gradient of bovine serum albumin and fractionated into five subpopulations. Cells from each subpopulation were assayed for spontaneous cytotoxic activity against K562 tumor cells. Immediately following fractionation, monocytes were not cytotoxic. Following incubation for at least 48 hr, monocytes from two layers of the gradients clearly exhibited greater spontaneous cytotoxic activity than all others. The degree of cytotoxicity expressed by cells of these layers was enhanced by the addition of indomethacin and inhibited by prostaglandin E2 (PGE2). Monocytes acquiring spontaneous cytotoxicity did not secrete measurable levels of PGE2 and had increased levels of purine nucleoside phosphorylase after 72 hs of culture in vitro. Surface markers HNK-1 and Mac-1 normally associated with cytotoxic function, were detected on these cells by indirect immunofluorescence at isolation and after culture. The fraction with the greatest cytotoxic activity showed an increase in the proportion of cells displaying reactivity to HNK-1 after culture compared to initial isolation.

Persistently altered T cell immunity in high school students with the congenital rubella syndrome and profound hearing loss.

Because there are frequent progressive and autoimmune complications in children born with the congenital rubella syndrome, we evaluated immunoregulation in eight profoundly deaf adolescents with congenital rubella syndrome who lived in a state school. Serum antiviral antibodies, expressions of peripheral lymphocyte epitopes and serum soluble interleukin 2 receptor (IL-2R) content were compared with those of 16 classmates with profound hearing loss of unknown cause and of 24 age-matched, hearing students from this area. Both deaf groups showed activated but impaired T lymphocyte function compared with normals. Rubella virus alteration of T cell function was suggested in congenital rubella syndrome students by elevated numbers of both CD4+ helper and CD25+ IL-2R cells with unusually low released soluble IL-2R content. In contrast in deaf classmates elevated CD25+ and CD16+ natural killer cell groups and soluble IL-2R content with low numbers of CD4+ helper cells and CD4+ populations were of unknown etiology. Defective immunoregulation of the congenitally deaf to pathogens inherent in their environment may lead to autoimmune and other complications.

Role of the thymus in histiocytosis-X.

An increasing number of reports have demonstrated that patients with histiocytosis-X show significant morphologic changes in the thymus gland. The changes include severe dysplasia, dysmorphia, and severe nonspecific involution. These findings are present in all children with histiocytosis-X who die, but can be found even when the disease is limited to one bone and is not fatal.

Evidence of persistent viral infection in Menière's disease.

Aspects of humoral and cell-mediated immunity that might characterize the continuation of symptoms in 25 patients with chronic Meniere's disease were examined. We found significant elevations of both humoral and cellular immune responses to viral antigens of herpes simplex I, varicella-zoster, rubella, and cytomegalovirus. Serum immunoglobulins were quantitatively abnormal in 24 of 25 patients, without a consistent pattern. These immune responses may be linked to persistent viral infection in chronic Meniere's disease.

Quantitation of glycosaminoglycans of rabbit lung during delayed-type hypersensitivity reactions and granuloma formation.

The specificity and kinetics of hyaluronic acid (HA) accumulation in relation to other glycosaminoglycans (GASs) were determined in rabbit lungs during an allergic granulomatous response to BCG, an allergic nongranulomatous response to tuberculoprotein, and during a foreign-body granulomatous response to carrageenan. Hyaluronic acid was the only GAG detected in the lung lavage fluids. Hyaluronic acid occurred in the airways on day two of the allergic granulomatous response, but its presence in the airway did not correlate with ensuing granuloma formation in the parenchyma. Generalized increases in GAG of the parenchyma also peaked on day two of the DTH responses. Generalized increases in GAG peaked on day five during the foreign-body granulomatous response to carrageenan. A persistently elevated level of HA in the lung tissue correlated with granuloma formation but not with the intensity of the response.

The piebald-lethal murine strain: investigation of the cause of early death.

Forty-eight piebald-lethal (PL) mice with distal aganglionosis and 42 normal littermates (LM) were studied to determine the cause of early death. PL mice were noted to be smaller than their LM and to have normal albumin and immunoglobulin levels for the first 30 days of age. As PL mice aged, a significant decline in albumin with a concomitant rise in immunoglobulin levels was demonstratable. Systemic sepsis with enteric organisms was found in 10% of sacrificed PL mice and in 38% of spontaneously dying PL animals. Histologic examination of PL aganglionic and ganglionic colon demonstrated no evidence of enterocolitis. Ganglionic colon of PL mice contained a flattened, thinned mucosa. The early death of PL mice is related to generalized debilitation from prolonged distal colonic obstruction resulting in a decrease in immunologic integrity and an increased susceptibility to sepsis.

Additional evidence for the role of hyaluronic acid in the macrophage disappearance reaction.

A slight macrophage disappearance reaction was elicited in C3H/Hej mice, a low responder strain. Low levels of hyaluronic acid accumulated in the peritoneal fluid during the disappearance reaction, in keeping with the low response. The macrophage disappearance reaction was not simulated by the intraperitoneal injection of large amounts of hyaluronate into nonsensitized C3H/Hej mice. This suggested that these macrophages were refractory to hyaluronic acid. Subsequent experiments demonstrated that low concentrations of hyaluronic acid did not agglutinate these cells, in vitro, nor inhibit their migration to the extent obtained with macrophages from the responder mice (C57BL/6 strain), thus providing evidence that the low responder mice had at least two defects with respect to the disappearance reaction: 1) they accumulated less hyaluronic acid in their peritoneal fluid, and 2) their macrophages were refractory towards hyaluronic acid. Generalized defects in membranes of various cell types of C3H/Hej mice, involving receptors for lymphokines in the case of mesothelial cells, or hyaluronic acid receptors on macrophages may serve as the central defect and explain the above observations.

Suppressor-cell dysfunction in children with histiocytosis-X.

Suppressor-cell activity of peripheral blood mononuclear cells were examined and lymphocyte subsets analyzed in children with histiocytosis-X and in healthy, age-matched subjects. Suppressor-cell function was assessed by two methods, the indomethacin stimulation of mitogen-activated cultures and the concanavalin A-inducible suppressor-cell assay. The results of these two assays indicate that children with active disease have significantly decreased suppressor-cell activity. Additionally, the percentage and absolute number of OKT8+ lymphocytes were decreased in children with active disease. Suppressor-cell activity and lymphocyte subsets returned to normal, baseline levels with disease remission. This study documents for the first time suppressor-cell dysfunction and supports previous investigations in which suppressor T lymphocytes are deficient in children with active disease. These findings may explain certain clinical manifestations seen in children with histiocytosis-X.

Immune alterations associated with T lymphocyte activation and regulation in retinitis pigmentosa patients.

Altered immunoregulation, suggested previously in the heredo-familial retinal degeneration, retinitis pigmentosa (RP), led us to examine cell-mediated immune responses in 58 RP patients who expressed either recessive or dominant inheritance. Increased absolute numbers of activated T-cells, quantitated by flow cytometry using the Ta1 epitope, were found in the peripheral blood of 60% of these RP patients. The unusual finding was equally distributed between dominant (16 of 25) and recessive (19 of 33) types of RP, suggesting an immune process present in both patterns of heredity. Additional altered lymphocyte immunoregulation was suggested in all RP by lymphocyte responses to stimulants modified by indomethacin or histamine in vitro. Although no clear association could be found between Tal expression and demographic factors including age, sex, years with RP symptoms, or percentage of life with RP disease, the significantly altered immunoregulatory responses in RP may be related to the pathogenesis of RP.

Circulating cytotoxic immune components in dominant Charcot-Marie-Tooth syndrome.

Activated T cells, measured repeatedly in the demyelinating peripheral neuropathy, Charcot-Marie-Tooth syndrome (CMT; hereditary motor sensory neuropathy), might participate in myelin loss by a destructive inflammatory autoimmune process. To explore this possibility, plasma proportions of hydroxyleukotrienes, their fatty acid precursor, arachidonic acid, and lymphocyte epitopes associated with immune cell activation expression were measured in 18 adults with dominant, Type I CMT. Compared to age-matched normal controls, CMT I patients showed eicosanoid-linked immunoactivation by an elevated content of 12-hydroxy-eicosatetraenoic acid (12-HETE) in parallel with a decreased plasma percentage of its fatty acid precursor, arachidonic acid. CMT patients also had increased numbers of peripheral lymphocytes expressing activation-related epitopes, CD25+, CD26+, CD4+, and CD4/CD45RO+ primed memory cells, with enhanced CD8+ cytotoxic cells and soluble CD8 protein content. Therefore, endogenously stimulated CMT I lymphocytes include functional cytotoxic cells which appear to deplete the plasma fatty acid precursor of prostenoid agents during the secretion of potentially destructive cytokines.

Participation of hyaluronic acid in the macrophage disappearance reaction.

Hyaluronic acid accumulates in the peritoneal cavity of mice during the macrophage disapperance reaction. Macrophages reappear in the exudates during the depletion of hyaluronate. The disappearance reaction is simulated by the intraperitional (i.p.) injection of hyaluronic acid into normal mice. The disapperance reaction largely is inhibited by the simultaneous injection (i.p.) of specific hyaluronidase with the challenge dose of BCG. We propose that hyaluronic acid makes a major contribution to the macrophage disappearance reaction.

Selective suppression of granuloma formation and delayed hypersensitivity in rabbits.

The relationship between dermal delayed hypersensitivity (DH) and granulomatous hypersensitivity was studied in rabbits sensitized with killed mycobacteria. Specific antigen challenge of sensitized animals resulted in extensive pulmonary granulomatous inflammation and induced suppression of both dermal DH and dermal granuloma formation. Whereas suppression of DH was concomitant with pulmonary granuloma formation, as is the case in a number of granulomatous diseases, a causal relationship between the two did not exist. Both DH and dermal granulomatous hypersensitivity were significantly suppressed whether or not the antigen challenge was of a granulomagenic (particulate) or nongranulomagenic (soluble) form. The data presented indicate that granulomatous hypersensitivity and DH are selectively suppressed with regard to different anatomical sites.

Effect of neonatal cerebrospinal fluid on monocyte chemotaxis.

We have reported in our previous study that macrophages constitute 58% of the total number of white cells in noninfected neonatal cerebrospinal fluid (CSF). CSF macrophages are probably derived from blood monocytes. To explain their predominance, we compared the chemotactic response of monocytes from cord and adult blood to neonatal (NCSF) and non-neonatal spinal fluids (NNCSF). Zymosan-activated serum (ZAS) was used as a positive control. Significance was accepted as p less than or equal to 0.05. We observed that both random migratory (RM) activity and chemotactic responses (CM) of cord monocytes to neonatal CSF and ZAS were greater than those of monocytes obtained from adult blood, resulting in a significant increase in the chemotactic differential (CD). Chemotactic responses of neonatal monocytes were significantly greater in the presence of neonatal CSF as compared to CSF from older children as indicated by CD. The CD of neonatal monocytes to neonatal CSF was greater than that of neonatal neutrophils. These findings are consistent with the presence of specific monocyte chemotaxins in noninfected neonatal CSF.

Gm and Km allotypes in Charcot-Marie-Tooth disease.

Gm and Km immunoglobulin allotypes were studied in 46 Caucasian patients with Charcot-Marie-Tooth disease (CMT). No significant association of Gm and Km phenotypes with CMT was found. Family studies revealed that Gm haplotypes and CMT were not inherited together, thus arguing against the involvement of immunoglobulin allotypes in CMT.