Molecular and functional characterization of cynomolgus monkey IgG subclasses.

Studies for vaccine and human therapeutic Ab development in cynomolgus monkeys (cynos) are influenced by immune responses, with Ab responses playing a significant role in efficacy and immunogenicity. Understanding the nature of cyno humoral immune responses and characterizing the predominant cyno IgG types produced and the Fc-FcγR interactions could provide insight into the immunomodulatory effects of vaccines. Anti-drug Ab responses against human IgG therapeutic candidates in cynos may affect efficacy and safety assessments because of the formation of immune complexes. There is, however, limited information on the structure and function of cyno IgG subclasses and how they compare with human IgG subclasses in Fc-dependent effector functions. To analyze the functional nature of cyno IgG subclasses, we cloned four cyno IgG C regions by using their sequence similarity to other primate IgGs. The four clones, cyno (cy)IGG1, cyIGG2, cyIGG3, cyIGG4, were then used to construct chimeric Abs. The sequence features of cyno IgG subclasses were compared with those of rhesus monkey and human IgG. Our data show that rhesus monkey and cyno IgG C regions are generally highly conserved, with differences in the hinge and hinge-proximal CH2 regions. Fc-dependent effector functions of cyno IgG subclasses were assessed in vitro with a variety of binding and functional assays. Our findings demonstrate distinctive functional properties of cyno IgG subclasses. It is notable that human IgG1 was less potent than cyno IgG1 in cyno FcγR binding and effector functions, with the differences emphasizing the need to carefully interpret preclinical data obtained with human IgG1 therapeutics.

Safety of Benazepril in 400 Azotemic and 110 Non-Azotemic Client-Owned Cats (2001-2012).

This retrospective study examined cats after initiation of benazepril therapy to determine the frequency of systemic hypotension or elevations in serum creatinine and/or potassium. Medical records review identified azotemic and non-azotemic cats prescribed benazepril. Blood pressure was recorded at the first available time after initiation of therapy. No cats experienced documented systolic systemic hypotension (<90 mmHg). Serum creatinine, and potassium when available, were recorded at baseline and in time windows after initiation of treatment: 1-30 days and 31-60 days. Blood chemistry results were screened for hyperkalemia (≥6.0 mEq/L). During the first 2 mo after starting benazepril therapy, there was a low incidence (3.7%) and clinically insignificant magnitude of hyperkalemia. Serum creatinine increases of greater than 30% from baseline were noted. This change was found in 11.0% of cats during the first 30 days of therapy and in 13.7% of cats from days 31-60 after initiation of therapy. The long-term survival of the cats that had >30% increases in creatinine from baseline was not statistically different from the survival of those that did not experience these increases, which suggests this finding may not be a reason to discontinue therapy. Benazepril appeared safe in a heterogeneous population of cats.

Prevalence and underlying causes of histologic abnormalities in cats suspected to have chronic small bowel disease: 300 cases (2008-2013).

OBJECTIVE: To determine prevalence of histologic abnormalities in cats suspected, on the basis of compatible clinical signs and ultrasonographic findings, to have chronic small bowel disease; identify the most common underlying causes in affected cats; and compare methods for differentiating among the various causes of chronic small bowel disease. DESIGN: Retrospective case series. ANIMALS: 300 client-owned domestic cats suspected to have chronic small bowel disease. PROCEDURES: Medical records were reviewed to identify cats evaluated because of chronic vomiting, chronic small bowel diarrhea, or weight loss that also had ultrasonographic evidence of thickening of the small intestine. Cats were included in the study if full-thickness biopsy specimens had been obtained from ≥ 3 locations of the small intestine by means of laparotomy and biopsy specimens had been examined by means of histologic evaluation and, when necessary to obtain a diagnosis, immunohistochemical analysis and a PCR assay for antigen receptor rearrangement. RESULTS: Chronic small bowel disease was diagnosed in 288 of the 300 (96%) cats. The most common diagnoses were chronic enteritis (n = 150) and intestinal lymphoma (124). CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that a high percentage of cats with clinical signs of chronic small bowel disease and ultrasonographic evidence of thickening of the small intestine had histologic abnormalities. Furthermore, full-thickness biopsy specimens were useful in differentiating between intestinal lymphoma and chronic enteritis, but such differentiation was not possible with ultrasonography or clinicopathologic testing alone.

Dickkopf-1 regulates bone formation in young growing rodents and upon traumatic injury.

The physiological role of Dickkopf-1 (Dkk1) during postnatal bone growth in rodents and in adult rodents was examined utilizing an antibody to Dkk1 (Dkk1-Ab) that blocked Dkk1 binding to both low density lipoprotein receptor-related protein 6 (LRP6) and Kremen2, thereby preventing the Wnt inhibitory activity of Dkk1. Treatment of growing mice and rats with Dkk1-Ab resulted in a significant increase in bone mineral density because of increased bone formation. In contrast, treatment of adult ovariectomized rats did not appreciably impact bone, an effect that was associated with decreased Dkk1 expression in the serum and bone of older rats. Finally, we showed that Dkk1 plays a prominent role in adult bone by mediating fracture healing in adult rodents. These data suggest that, whereas Dkk1 significantly regulates bone formation in younger animals, its role in older animals is limited to pathologies that lead to the induction of Dkk1 expression in bone and/or serum, such as traumatic injury.