University of California, Davis (UCD) line 200 White Leghorn Chickens spontaneously develop a syndrome that has many analogous features to human progressive systemic sclerosis. This syndrome is characterized by progressive involution of comb, dermal fibrosis, and distal polyarthritis. These three features occur within 6 wk after hatching, and are accompanied by a 40% mortality as a result of vaso-occlusive disease, with development of secondary infection of peripheral gangrenous lesions. Birds that survive greater than 2 mo after hatching progressively develop fibrosis of the esophagous and mononuclear infiltration of heart and kidney, with prominent occlusion of small and medium sized blood vessels. In addition, line 200 chickens develop rheumatoid factors, antinuclear antibodies, and antibodies to collagen, but do not have antibodies to thymocytes, DNA, or extractable nuclear antigens. Moreover, antinuclear antibodies when studied using HEp-2 cells as substrate demonstrate predominantly a speckled pattern. This syndrome of line 200 chickens is not detectable in F1 crosses to several UCD inbred lines. F1 X parental line BC1 backcrosses have an approximately 50% incidence of disease, suggesting that this syndrome is inherited as autosomal recessive. However, only 4% of F2 generation birds show abnormal symptoms, suggesting the presence of modifying genes. There is no appearance of IgG deposition, as determined by immunofluorescence, in either skin, blood vessels, esophagus, or heart. However, approximately 20% of chickens have a glomerulonephritis; this feature appears to be a terminal event and does not appear clinically significant. Although this syndrome of line 200 chickens has several features that are in sharp distinction to human scleroderma, the presence of common immunologic and pathologic denominators suggest that this spontaneous disease may be an appropriate model to develop a better understanding of autoimmune connective tissue diseases.
Abstract. University of California, Davis line 200 White Leghorn chickens develop an inherited progressive fibrotic disease that includes the appearance of antinuclear antibodies (ANA). To further characterize these ANA, serial aged line 200 birds were studied. Greater than 50% of line 200 birds develop antinuclear and anticytoplasmic antibodies; fluorescent staining patterns included cytoplasmic spider web, most prevalent at 1 mo of age, and fine speckled patterns, characteristic of chickens 6 mo and older. By enzyme-linked immunosorbent assay, 40.4% of line 200 birds were found to have antibodies to single-stranded DNA (ssDNA). In contrast, antibodies to histones, RNA, or poly A * poly U were not detected. Precipitating antibodies to saline extracts from chicken liver were noted in 33.3% ofline 200 birds. Saline extracts from turkey, pheasant, and partridge liver but not rat, rabbit, or mouse tissues were also positive in immunodiffusion testing with these line 200 birds. The antigenicity of chicken liver extracts was sensitive to pronase, protease K, and pH variations >10 and <5; however, they were resistant to trypsin, DNase, RNase, and incubation at 370C and 560C for 1 h. Cell fractionation in conjunction with column chromatographic techniques revealed that several protein antigens with apparent molecular weights in the range of 62,000-290,000 were present in cytoplasm but not in isolated nuclei. Line 200 sera were not reactive against nuclear ribonucleoprotein, Sm, Scl-70, or SS-B/La antigens. Thus, line 200 chickens develop antinuclear and anticytoplasmic antibodies at an
Rhesus monkey infants fed a marginally zinc-deficient diet (4 ppm) from conception through 12 mo of postnatal life were monitored for changes in hematological, biochemical, and immunological parameters. These zinc-deprived (ZD) infants were compared to control infants whose mothers were fed a zinc-replete (100 ppm) diet either ad libitum (AL) or pair-fed (PF) throughout gestation and lactation. Blast transformation of peripheral blood lymphocytes to phytohemagglutinin (PHA-P), concanavalin A (Con A), and pokeweed mitogen (PWM), was dramatically depressed in the zinc-deficient (ZD) group. Similarly, ZD infants had reduced polymorphonuclear leukocyte function as measured by chemotaxis to endotoxin-activated plasma and phagocytosis of Candida albicans. Levels of serum IgM were significantly altered in zinc-deficient infants compared to controls. Serum concentrations of IgG and IgA were similar in zinc-deficient and control infants. ZD infants also manifested a hypochromic microcytic anemia at one month of age, reduced activity of the zinc metalloenzyme alkaline phosphatase, and lower activity of SGPT.
To assess long-term effects of marginal zinc deprivation on pregnancy, adult non-pregnant female rhesus monkeys were fed diets containing 100 or 4 ppm zinc for 1 yr. then mated; effects on pregnancy and its outcome are under study. During this year, food intake was not depressed in zinc-deprived (ZD) monkeys, and there were relatively few effects on biochemical or hematological indices. By the end of the year, plasma zinc concentration was somewhat lower in ZD monkeys than in controls. Several immune variables, including serum IgM and IgG levels and polymorphonuclear leukocyte (PMN) function, were depressed in the ZD group, changes closely reflecting circannual fluctuations in plasma zinc levels in both diet groups. Endotoxin-activated plasma from ZD monkeys had less potential to promote chemotaxis than that from control monkeys, suggesting that defective PMN function may relate to a plasma effect. Marginal zinc deprivation may thus influence immune function before other variables are affected.
The "LipoGen RheumaStrip ANA Profile" test method (LipoGen, Inc.) is a new assay format for autoantibody detection in which recombinant autoantigens are used. This enzyme immunoassay, in test-strip format, detects antibodies to autoantigens U1-ribonucleoprotein (U1-RNP), Sm, SS-A/Ro, SS-B/La, and to native DNA (nDNA). We evaluated 200 antinuclear antibody (ANA)-positive and 100 ANA-negative sera for the presence of antibodies to U1-RNP, Sm, SS-A/Ro, SS-B/La, and nDNA by the new test-strip procedure. These data correlated well with those obtained with either Ouchterlony double immunodiffusion for U1-RNP, Sm, SS-A/Ro, and SS-B/La or with Crithidia luciliae indirect immunofluorescence for anti-nDNA. Assay sensitivity and assay specificity of the ANA Profile method as compared with those of established procedures were respectively as follows: 89.8% and 98.8% for U1-RNP, 86.4% and 95.3% for Sm, 97.9% and 89.3% for SS-A/Ro, 98.3% and 86.3% for SS-B/La, and 97.5% and 93.1% for nDNA. Agreement between the ANA Profile test and these other test methodologies ranged from 88.7% for the SS-B/La test to 97.3% for the U1-RNP test. This new test procedure substantially decreases the time and effort required to perform these assays. Total hands-on time and overall assay time were decreased by 72% and 97%, respectively.
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