Dendritic cell therapy in composite tissue allotransplantation.

Dendritic cells (DCs) are bone marrow-derived, professional antigen-presenting cells, with inherent tolerogenic function. The ability of immature or maturation-resistant DCs to regulate alloantigen-specific T-cell responses and to promote tolerance induction has been well demonstrated in organ and bone marrow transplantation. Recent data suggest that DCs can also promote long-term survival of composite tissue allografts in the absence of continued immunosuppressive drug therapy.

Specimen Collection for Translational Studies in Hidradenitis Suppurativa.

Hidradenitis suppurativa (HS) is a chronic inflammatory disorder characterized by painful nodules, sinus tracts, and scars occurring predominantly in intertriginous regions. The prevalence of HS is currently 0.053-4%, with a predominance in African-American women and has been linked to low socioeconomic status. The majority of the reported literature is  retrospective, population based, epidemiologic studies. In this regard, there is a need to establish a repository of biospecimens, which represent appropriate gender and racial demographics amongst HS patients. These efforts will diminish knowledge gaps in understanding the disease pathophysiology. Hence, we sought to outline a step-by-step protocol detailing how we established our HS biobank to facilitate the formation of other HS tissue banks. Equipping researchers with carefully detailed processes for collection of HS specimens would accelerate the accumulation of well-organized human biological material. Over time, the scientific community will have access to a broad range of HS tissue biospecimens, ultimately leading to more rigorous basic and translational research. Moreover, an improved understanding of the pathophysiology is necessary for the discovery of novel therapies for this debilitating disease. We aim to provide high impact translational research methodology for cutaneous biology research and foster multidisciplinary collaboration and advancement of our understanding of cutaneous diseases.

The Pittsburgh study of insulin-dependent diabetes mellitus. Risk for diabetes among relatives of IDDM.

AN analysis has been made of the family histories of a survey of 1280 cases of IDDM entering Children's Hospital of Pittsburgh between December 31, 1964 and January 1, 1981, discharged on insulin and initial age of onset under 17 yr. Family histories revealed an increased occurrence of IDDM among relatives in the affected families. The risk to siblings was estimated by age-corrected proband exclusion (3.3%) by age 20 and by the Li-Mantel segregation ratio estimator (6.0%). The comparison of these risk measures is discussed. The occurrence of IDDM among the parents is 2.6% and of NIDDM among the parents is 2.4%. A comparison of risk to relatives (parents, sibs, uncles, half-sibs) observed in the Pittsburgh Study to those of six other studies reveal essentially equivalent rates. There is no increased risk to siblings of a diabetic who had an early age of onset. There is an increased risk to siblings of a diabetic (10.5%) in families where at least one parent has insulin-dependent diabetes mellitus (IDDM) and also an increased risk to siblings of a diabetic (8.8%) when at least one parent has non-insulin-dependent diabetes (NIDDM). The average age of onset for second cases in a family is significantly older than age of onset in single case families.

Immunological surveillance in a pseudorabies quarantined herd using gilts and their progeny as sentinels.

Specific pathogen free gilts and their progeny were evaluated to use as sentinels in a pseudorabies virus (PRV) infected herd by immunologically monitoring for PRV seroconversions. Time intervals targeted were pre- and post-PRV vaccinations, herd exposure, and farrowing to finishing. Post-PRV vaccinations, gilts showed low PRV lymphocyte stimulation and humoral responses. Following herd exposure, control gilts PRV seroconverted and PRV vaccinated gilts increased (2 to 4 times) in virus neutralization (VN) titers. Sixty-seven percent (4/6) of the progeny from a control gilt were PRV seropositive at finishing. Progeny from PRV vaccinated gilts were depleted of passive immunity by week 7, and were seronegative until week 9. At finishing 47% (14/30) of them were PRV seropositive indicating exposure to PRV. The VN test was not sensitive enough to detect weak positive serums, noted as positives by latex agglutination (LA) test, ELISA, and Western blots. The gilts and progeny detected PRV, respectively, in the herd housing quarters and in the farrow to finish facilities. A strategy for future sentinel experimental surveillances using primarily the LA test is proposed.

Immunity induced in rats vaccinated with toxoid prepared from heat-labile toxin produced by Pasteurella multocida serogroup D.

Rats were vaccinated with a toxoid (D-toxoid) prepared from purified heat-labile toxin (D-toxin) produced by Pasteurella multocida serogroup D. Vaccination of rats with D-toxoid prevented death and other effects of D-toxin (hepatic necrosis, development of elevated leukocyte counts, lymphopenia, neutrophilia, and elevated complement titers) that occurred in phosphate buffered saline (PBS)-vaccinated control rats.

Breed susceptibility to ovine progressive pneumonia (maedi/visna) virus.

In this retrospective study of breed differences in susceptibility to disease caused by ovine progressive pneumonia (OPP) virus, 29 Border Leicester sheep were compared with 46 Columbia sheep. As judged by frequency and severity of clinical signs and lesions attributable to the infection, Border Leicester sheep were markedly more susceptible than Columbia sheep and experimentally infected sheep were slightly more susceptible than naturally infected sheep. Differences in susceptibility to infection by the virus were not determined.

Effects of adjuvant-augmented germling vaccines in turkey poults challenged with Aspergillus fumigatus.

Turkey poults were vaccinated with combinations of two different germling preparations and three adjuvants (N-acetylmuranyl-L-alanyl-D-isoglutamine, Pasteurella multocida lipopolysaccharide [LPS], and avridine) at 1 and 2 weeks of age, and their immunity was challenged by sublethal exposure to aerosols of Aspergillus fumigatus conidia at 1 month of age. Fewer turkeys in the groups given vaccines prepared from germlings grown on Dorset's and Henley's medium (D&H) had organisms in lung tissue at 2 weeks after challenge exposure as compared with those vaccinated with germling grown on neopeptone dialysate (Neo). The LPS of P. multocida appeared to be the most efficacious of the adjuvants in the D&H vaccine group, as A. fumigatus was isolated from only one of eight turkeys in this group; the number of organisms per gram of lung tissue was low compared with other vaccine groups at 2 weeks after challenge exposure; and poults given D&H vaccine with LPS as adjuvant had less-severe lung lesions than other groups. These differences in lung lesions were more marked at 2 weeks than at 8 weeks after challenge exposure. The only difference among other parameters in the vaccinated turkeys was lower heterophil counts in the turkeys given D&H-prepared vaccines than in unvaccinated controls. This was probably due to less-severe infections resulting from protective effects of these vaccines.

Development and evaluation of an enzyme-linked immunosorbent assay for bovine antibody to Pasteurella haemolytica: constructing an enzyme-linked immunosorbent assay titer.

A 2-stage strategy was developed and evaluated for estimating serum antibody titer by use of ELISA and a series of dilutions. In stage 1, the linear response region and least-square estimate of the assay line slope were established from 9-point dilution assays. Provided that the reading was within the linear response region, this information was used in the stage-2 estimation of titer from a single absorbance reading. Operationally, 2 fixed dilutions were selected, one suitably low and one suitably high, to provide at least one reading within the linear region. The procedure should save considerable time when a large number of assays are to be performed. Stage 1 required approximately twenty 9-point assays, but all subsequent assays required only 2 fixed dilutions.

Frequency of association of noncytopathic bovine viral diarrhea virus with mononuclear leukocytes from persistently infected cattle.

All mononuclear leukocytes and T lymphocyte-enriched and B lymphocyte-enriched subpopulations of mononuclear leukocytes collected from 8 cows persistently infected with 1 of 3 isolates of noncytopathic bovine viral diarrhea virus were tested for association with virus. For all persistently infected cows, approximately 4.4% of all mononuclear leukocytes, 5.4% of T lymphocyte-enriched, and 2.1% of B lymphocyte-enriched subpopulations of mononuclear leukocytes were associated with virus. Differences between leukocyte populations in percentages of leukocytes associated with virus were real (P less than 0.05). Among virus isolates, significant differences in percentages of leukocytes associated with virus were not detected.

Prevalence estimation from pooled samples.

Under certain conditions, prevalence can be estimated by testing samples from individual members or by pooling samples from members into a group and by testing the sample from the group as a single unit. Pooled tests are more accurate than individual tests when P is less than 10%. Optimal group size is 1.6/P. Efficiency decreases slowly with suboptimal size and rapidly with overly large size.

Development and evaluation of an enzyme-linked immunosorbent assay for bovine antibody (IgG) to Pasteurella haemolytica.

The sensitivity of an indirect enzyme-linked immunosorbent assay (ELISA) for bovine IgG serum antibody to Pasteurella haemolytica was compared with that of an indirect hemagglutination (IHA) test. Pasteurella haemolytica serotypes were grown in a chemically defined cell culture medium, and soluble antigens released into the growth medium were used in the ELISA and IHA test. An ELISA with serotype-1 antigen consistently detected antibody in sera that were positive by IHA test (correlation, 99%). Sera reacting with serotype-1 ELISA antigens also reacted with ELISA antigens prepared from other serotypes. Although ELISA titers averaged 5 log2 units higher than IHA titers, plots of titers determined by the 2 methods were approximately linear. Titer increases detected in paired serum samples by either test were similar. The ELISA was more sensitive than was the IHA in detecting colostral IgG antibody in serum of newborn calves. The ELISA uses a simple, stable antigen preparation and detects antibody to P haemolytica serotypes that commonly infect cattle.

Antiviral effectiveness of butylated hydroxytoluene against pseudorabies (Aujeszky's disease) virus in cell culture, mice, and swine.

Butylated hydroxytoluene (BHT) was evaluated for antiviral effectiveness on pseudorabies virus (PRV) in cell culture, mice, and swine. When relatively small amounts of BHT were mixed with PRV and incubated at 37 C for 30 or 60 minutes before inoculation into cell cultures, the cell cultures did not become infected with virus. The PRV was not infectious when the virus was treated with BHT and then inoculated intraperitoneally into mice, but was infectious when BHT and PRV were inoculated simultaneously or when BHT was inoculated either 30 or 60 minutes before PRV. Swine fed BHT-medicated feed for 10 days before they were intranasally exposed with virulent PRV did not have overt signs of pseudorabies, had a lower concentration of PRV in nasal mucus than did control swine, and had acceptable blood enzyme and cholesterol concentrations during the experiment. The BHT was detected in tissues of 2 swine after they were fed BHT-medicated feed for 10 days, and higher concentrations of BHT were detected in tissues of 3 swine given BHT feed for 29 days.

Decreased complement and bacteriostatic activities in the sera of cattle given single or multiple doses of aflatoxin.

Five steers given 1 dose of partially purified aflatoxin at concentrations sufficient to provide 0.2 mg to 0.8 mg of aflatoxin B1 equivalents/kg of body weight were compared with 4 steers given 14 daily doses of 0.25 mg of aflatoxin B1 equivalents/kg of body weight for complement activity and bacteriostasis. Complement activity was measured by hemolysis in gel, and bacteriostatic activity, by growth inhibition of Escherichia coli in liquid medium. In the single-dose group, complement activity and bacteriostatic activities decreased by 57 hours after dosing, and both returned to near base line by 168 hours. In the daily dose group, only bacteriostatic activity decreased, and the decrease persisted 2 weeks after the last dose of aflatoxin was given. Apparently, aflatoxin affects both complement-dependent and independent serum bacteriostatic activity.

Seroprevalence of ovine progressive pneumonia virus in sheep in the United States as assessed by analyses of voluntarily submitted samples.

Ovine progressive pneumonia (OPP) is a lentivirus-induced disease of sheep in the United States that is similar, if not identical, to maedi/visna in many other countries. Prevalence estimates of seropositivity to this virus in sheep in the United States have been confined to limited groups or flocks of sheep and have varied from 1 to 90%. In this study of detection of antibodies against OPP virus, we found a lower general prevalence of antibodies to OPP virus in sheep than was previously reported. Of 16,827 sheep from 29 states in the United States, 26% were seropositive and 48% of 164 flocks that were tested had 1 or more seropositive sheep. Seropositivity to OPP virus for sheep within special categories was determined, although nonrandom samples that were available may have biased the results. Within regions of the United States, prevalence was highest in the Rocky Mountain region at 49% and lowest in the northern Atlantic region at 9%. Seropositive sheep were not evenly distributed among flocks, but were clustered in a few flocks of sheep. A high number of flocks had no or few seropositive sheep. Prevalence increased with age from 4% at less than 1 year to a plateau of 34% at 4 years. Seropositivity was variable among breeds and was not associated with sex, wool class, or place of origin of ancestors.

Evaluation of field isolates of pseudorabies (Aujeszky's disease) virus as determined by restriction endonuclease analysis and hybridization.

Sixty-seven isolates of pseudorabies virus (PRV) from 13 states were cleaved with 4 restriction endonucleases (RE), and after electrophoresis in agarose, their banding patterns were photographed and evaluated. The deoxyribonucleic acid (DNA) cleavage fragments were designated into regions specified by molecular weight ranges based on lambda phage DNA as a size marker. The 67 PRV isolates were evaluated according to the total number of cleavage fragments, by the number of fragments within designated molecular weight regions, and finally, by the migration of fragments within regions. Four of the 67 PRV isolates (all 4 from California) did not have a 4.1 to 4.6 megadalton HinfI band, but hybridization of the HinfI digests with a 32P probe made with the 4.4 megadalton band hybridized with 2 lighter fragments, 2.5 and 1.9 megadaltons, respectively. The BamHI digests of DNA from some PRV isolates with submolar fragments were hybridized with 32P probes made with fragments from the submolar region and the BamHI E fragment. Both probes hybridized to the submolar region of PRV with BamHI submolar fragments, but only to the trimolar (E, F, and G) band of PRV without submolar fragments in the 4.1 to 7.5 BamHI megadalton region. Epidemiologic evidence was obtained which indicated that a Missouri strain of PRV was transferred to an Illinois swine herd by importation of feeder pigs from Missouri. The results indicate that there are numerous genomically different PRV currently in circulation in the United States and that the combination of RE analysis and DNA hybridization offers useful epidemiologic information to evaluate the various strains.

Complement, bacteriostatic, and enzymatic activities in sera from guinea pigs given aflatoxin and/or rubratoxin.

The relationship of serum complement activity and bacteriostatic activity was investigated in male guinea pigs given aflatoxin and/or rubratoxin. In experiment 1, guinea pigs were given 0.6 mg of aflatoxin/kg of body weight, PO, once. In experiment 2, guinea pigs were given 0.02 mg of aflatoxin/kg, PO, and/or 8 mg of rubratoxin, PO, 11 times. Aflatoxin (0.02 mg/kg) had no effect given alone, but potentiated the effect of rubratoxin. In both experiments, changes in complement activity were accompanied by similar but not always significant (P less than 0.05) changes in bacteriostatic activity of serum. Guinea pigs given 0.06 mg of aflatoxin/kg had significant (P less than 0.05) changes in complement titers and in serum alkaline phosphatase, alanine aminotransferase, and aspartate aminotransferase activities. Guinea pigs given repeated oral doses of aflatoxin and/or rubratoxin had changes in complement titers, bacteriostasis, and alkaline phosphatase and aspartate aminotransferase activities, but not in alanine aminotransferase activities. Significant differences were detected only when average values for all guinea pigs given rubratoxin or rubratoxin with aflatoxin were compared with average values for guinea pigs not given rubratoxin.

Serologic survey for transmissible gastroenteritis virus neutralizing antibodies in selected feral and domestic swine sera in the southern United States.

Serum samples collected from feral and domestic swine (Sus scrofa) in Florida and feral swine in Georgia and Texas were assayed by plaque reduction for their virus neutralizing (VN) antibodies against the porcine transmissible gastroenteritis virus (TGE). None of 560 samples collected from feral swine contained VN antibodies for TGE virus, but experimentally infected feral swine seroconverted. None of 665 samples from domestic swine contained TGE-VN antibodies. These results indicate feral swine are not a significant reservoir for TGE virus in southern states, but are capable of becoming infected and developing VN antibodies against TGE.

Prevalence and transmission of pseudorabies virus in an isolated population of feral swine.

Six hundred sixty-one feral swine (Sus scrofa) from Ossabaw Island, Georgia (USA) were captured, bled, and their sera tested for pseudorabies virus (PRV) antibody during a 6 yr period. Prevalence of seroconversion in females was somewhat higher than in males (10% versus 7%), but the difference was not statistically significant. Adults had a significantly higher prevalence than juveniles (29% versus 1%). An important finding in this study was that seroconversion occurred primarily in the adult feral swine.

Prevalence of Mycobacterium paratuberculosis in ileocecal lymph nodes of cattle culled in the United States.

During 1983 and 1984, ileocecal lymph node specimens were obtained from clinically normal cattle at 76 US Department of Agriculture-inspected cull cattle slaughterhouses in 32 states and Puerto Rico. Mycobacterium paratuberculosis was isolated from the lymph nodes of 119 of 7,540 cattle. The prevalence of bovine paratuberculosis was 1.6% overall, with 2.9% in dairy culls and 0.8% in beef culls. The prevalence for females and males did not appear to differ significantly.