Preventing the development of SLE: identifying risk factors and proposing pathways for clinical care.

Although challenging, developing evidence-based approaches to an early and accurate diagnosis of systemic lupus erythematosus is a key approach to preventing disease and lupus-associated morbidity and mortality. Advances in our understanding of preclinical and incomplete lupus erythematosus have enabled the identification of risk factors that may predict disease and the development of potential strategies aimed at primary prevention. Emerging data support the notion that there is a temporal disease progression from initial asymptomatic autoimmunity (preclinical lupus) through early clinical features of the disease (incomplete lupus erythematosus) to finally becoming fully classifiable systemic lupus erythematosus (complete lupus erythematosus). Here, we review the demographic, clinical, biomarker as well as genetic and environmental features that are reported to increase the risk of disease progression. Based on these risk factors, we propose a clinical care pathway for patients with early disease. We envisage that such a pathway, through early identification of disease, may improve patient outcomes, while reducing health care costs.

Differences in immunocontraceptive responses in white-tailed deer (Odocoileus virginianus) and goats (Capra hircus).

Thirteen deer (Odocoileus virginianus) and three goats (Capra hircus) were given an initial immunization of 300 microg porcine zona pellucida (PZP) combined with 50 mg synthetic trehalose dicorynomycolate ml(-1) (STDCM) in drakeol i.m. in the semitendinosus and semimembranosus muscles. This immunization was followed by two booster injections of 300 microg PZP, 2 weeks apart. The vaccinations were made from the same purified batch of PZP and lot of adjuvant. All the doses were made at the same time and injected on the same day. The immune response was quantified by measuring serum anti-PZP IgG antibody concentration by ELISA. The results showed that the mean serum IgG concentration of the deer increased from 0.000 +/- 0.003 to 0.083 +/- 0.023 absorbance units, whereas in the goats the mean IgG concentration increase was from 0.044 +/- 0.019 to 1.245 +/- 0.774 absorbance units. Both the goats and deer showed a significant increase in IgG concentration (P < 0.05) after the final booster injection compared with preimmune serum. The final goat IgG concentration was significantly (P < 0.05) higher than that of the deer. If PZP and Freund's adjuvant are administered to deer, immunocontraception is achieved, but the IgG concentration in the deer in this study did not appear to be compatible with immunocontraception.

Immunolocalization of zona pellucida antigens in the ovarian follicle of dogs, cats, horses and elephants.

A comparative evaluation of the location of immunoreactive porcine zona pellucida (pZP) glycoproteins was performed with polyclonal rabbit anti-pZP antibodies on ovarian sections of the dog, cat, horse, and elephant. For this, formalin (light microscopy) and glutaraldehyde (transmission electron microscopy [TEM]) fixed ovarian sections were incubated with antibodies raised against highly purified pZP. Staining patterns were determined with diaminobenzidine (DAB) at the light level. The dog ZP had a distinct staining distribution that is characterized by intense staining around the periphery of the ZP and the oolemma and less dense staining throughout the width of the ZP. In dog follicles that contained multiple oocytes, there were oocytes of identical and dissimilar stages. Cat ovarian sections showed uniform staining of the ZP. Horse results showed uniform staining of ZP and ooplasm, and granulosa cells (GC). Elephant sections showed staining of the ZP with dense staining at the oolemma, as well as staining of the ooplasm. In all species the staining of the ZP was not evident until GC differentiation. In all cases there was no staining of ovarian tissue with control normal rabbit serum. Specific staining patterns of ZP were evaluated by TEM and immunogold staining. The immunogold-linked anti-pZP antibodies stained the ZP matrix in all species. There was staining of ooplasm organelles suggesting that ZP secretion originates from the oocyte of the dog and cat. In addition, follicular and ZP measurements were taken that allowed accurate characterization of follicle stage. These findings suggest that in all four species the ZP is recognized by anti-pZP antibodies and there is also evidence to suggest the possible origins of ZP glycoproteins.

Evaluation of somatic and reproductive immunotoxic effects of the porcine zona pellucida vaccination.

Immunological, immunocytochemical and fertility analyses were performed to determine the potential toxic side effects of porcine zona pellucida (pZP) vaccinations on target animals, including horses and dogs. The study was designed to determine the effect of antibodies, raised against highly purified pZP, on somatic tissues. Immunocytochemical studies performed with fixed tissues showed that rabbit anti-pZP antiserum did not crossreact with brain, heart, lung, kidney, liver, bladder, stomach, small intestine, large intestine, muscle, skin, spleen, pancreas, or lymph node of either the dog or horse. To determine the effect or oral intake on nontarget animals, female rabbits were fed a contraceptive vaccine containing pZP glycoproteins and the synthetic trehalose dicorynomycolate in drakeol (S-TDCM) adjuvant. Enzyme-linked immunoadsorbent assay (LISA) analyses showed that rabbits fed with the adjuvanted pZP proteins did not develop circulating anti-pZP IgG antibodies that crossreacted with pZP. Furthermore, fertility studies performed on rabbits fed with adjuvanted pZP revealed no significant differences in the number of embryos or stage of the embryos produced between the treated and control animals. Results of these studies suggest that the pZP vaccine delivered to dogs or horses in field studies have no recognizable somatic tissue effects. Moreover, there were no side effects on nontarget animals should they eat the vaccine. This substantiates field trials results about the safety of the pZP immunocontraceptive vaccine.

Possible mechanisms of mammalian immunocontraception.

Ecological and conservation programs in ecosystems around the world have experienced varied success in population management. One of the greatest problems is that human expansion has led to the shrinking of wildlife habitat and, as a result, the overpopulation of many different species has occurred. The pressures exerted by the increased number of animals has caused environmental damage. The humane and practical control of these populations has solicited the scientific community to arrive at a safe, effective, and cost-efficient means of population control. Immunocontraception using zona pellucida antigens, specifically porcine zona pellucida (pZP), has become one of the most promising population control tools in the world today, with notable successes in horses and elephants. A conundrum has risen where pZP, a single vaccine, successfully induces an immunocontraceptive effect in multiple species of mammals. This review describes the most current data pertaining to the mammalian zona pellucida and immunocontraception, and from these studies, we suggest several potential mechanisms of immunocontraception.

Inducible and constitutive in vitro neutrophil chemokine expression by mammary epithelial and myoepithelial cells.

Previously, our laboratory showed that bovine and caprine mammary secretions are chemotactic and that chemoattractants found in these secretions are qualitatively different according to infection status and/or lactation stage. However, the cellular source of the chemoattractants has not been defined. In this study we used a modified Boyden chamber assay to examine the ability of previously established caprine mammary epithelial cell (CMEC) and myoepithelial cell (CMMyoEC) lines to produce chemoattractants for neutrophils. We found that CMEC culture supernatants, but not those of CMMyoEC cultures, induced in vitro neutrophil chemotaxis. Further characterization showed that chemotactic activity was produced when the cells underwent contact-induced differentiation. Neutrophil migration was chemotactic, not chemokinetic, and was augmented when the epithelial and myoepithelial cells were cocultured. Additionally, chemotactic activity was inducible by Staphylococcus aureus plus alpha-toxin, Escherichia coli, and interleukin-1beta (IL-1beta) in CMEC cultures. However, CMMyoEC cultures could not be induced to produce chemotactic activity. Anti-IL-8 antibody was able to block some constitutively produced chemotactic activity and chemotactic activity induced by IL-1beta and S. aureus plus alpha-toxin. These results indicate that epithelial cells may play a major role in producing chemoattractants, specifically IL-8, in the mammary gland.

Tumor infiltrating lymphocytes: CD8+ lymphocytes in canine transmissible venereal sarcomas at different stages of tumor growth.

This study utilized an in vivo tumor system (canine transmissible venereal sarcoma, CTVS) to assess wh correlation exists between tumor growth stage and phenotype of infiltrating lymphocytes. Additionally, the ability of CTVS cells to produce chemoattractants for canine lymphocytes was assessed. The CD8 subset of tumor infiltrating lymphocytes (TIL) during progressive growth, steady-state (no growth), and regression of the CTVS was determined. During progressive growth, the proportion of TIL expressing CD8 was significantly lower than that from regressing tumors (P < .001) and steady-state tumors (P < .01). Additionally, CTVS cell culture, CTVS spheroid cell culture, and CTVS spheroid/canine lymphocyte co-culture supernatants were tested for chemotactic activity for canine lymphocytes. CTVS and CTVS spheroid/canine lymphocyte co-culture supernatants both had significant chemotactic activity. Conversely, CTVS spheroid culture supernatants were negative for chemotactic activity for canine lymphocytes. These results indicate a correlation exists between the CD8 subset of TIL and clinical stage of the tumor. Also, we have shown that CTVS cells in vitro produce soluble factors that cause chemotaxis of canine lymphocytes. An understanding of the role of TIL and the ability of the tumor to attract them will be of help in designing strategies for immunomodulatory therapies for solid tumors.

Chemotactic activities in nonmastitic and mastitic mammary secretions: presence of interleukin-8 in mastitic but not nonmastitic secretions.

Due to its association with low-quality milk and a decrease in milk production in bovines, mastitis is a major cause of economic loss. Additionally, mastitis can be harmful to suckling newborns and can cause damage to the mammary gland. In mastitic mammary secretions there is a substantial increase in somatic cells, specifically neutrophils. In this study we examined the ability of mastitic and nonmastitic mammary secretions to cause in vitro neutrophil chemotaxis using a microchemotaxis assay. Also, the role of the inflammatory chemokine interleukin-8 (IL-8) in neutrophil recruitment during mastitis was addressed in these in vitro experiments. We found that both nonmastitic and mastitic mammary secretions were chemotactic, not chemokinetic, for neutrophils. The neutrophil chemotactic activity in mastitic, but not nonmastitic, mammary secretions was blocked by anti-IL-8 antibodies. Molecular mass separation of the active components showed that the chemotactic activity of the mastitic secretions was present in the 10-kDa-or-less fraction and was blocked by anti-IL-8 antibodies. These results indicate that IL-8 plays a major role in neutrophil recruitment during mastitis. An understanding of its role will be of help in designing strategies for immunomodulatory therapies for mastitis.

TCDD residues in fish and shellfish from U.S. waterways.

TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) in the edible portion of fish and shellfish from various U.S. waterways has been monitored since 1979. Analytical results for the period 1979-1994 are reported. Extracts obtained after column chromatographic and liquid chromatographic cleanup were examined by electron capture detection-gas chromatography (GC), and final quantitation and confirmation were performed by GC/mass spectrometry with multiple ion detection. Analyses of 1623 test samples indicated that TCDD residues in fish and shellfish were not widespread but rather were localized in areas near waste sites, chlorophenol manufacturers, and pulp and paper mills. Analytical results indicated that levels in aquatic species from these sites have been declining steadily. No TCDD (limit of detection and confirmation, 1-2 ppt) has been found in recent years in aquatic species from most Atlantic, Pacific, and Gulf of Mexico sites and Great Lakes other than Lake Ontario and Saginaw Bay (Lake Huron).