The anti-idiotypic antibody to chlamydial glycolipid exoantigen (GLXA) protects mice against genital infection with a human biovar of Chlamydia trachomatis.

Despite more than three decades of anti-chlamydial vaccine research and improved vaccine strategies with new technologies, no vaccine candidate has protected against heterologous challenge, nor at more than one site of infection. The majority of experimental anti-chlamydial vaccines to date have targeted the chlamydial major outer membrane protein (MOMP). Many MOMP-directed vaccine candidates have been highly immunogenic, but have failed to protect against infectious challenge. We have extended our previous studies of a different anti-chlamydial vaccine, a monoclonal anti-idiotypic antibody (anti-Id; mAb2) which is a molecular mimic of the chlamydial glycolipid exoantigen (GLXA). The present studies demonstrate that the mAb2 vaccine is protective in a murine genital infection model utilizing a human urogenital strain. After either mucosal (oral or intranasal) or systemic (subcutaneous) immunization with the poly (lactide) encapsulated-mAb2 to GLXA, C3H/HeJ mice were significantly protected against topical vaginal challenge with Chlamydia trachomatis (K serovar; UW-31). Reduced vaginal shedding of organism and genital tract inflammation were associated with GLXA-specific and/or anti-EB neutralizing serum antibody. Our results demonstrate that the anti-Id (mAb2) vaccine is protective against an additional human biovar of C. trachomatis in C3H/HeJ mice, which are allogeneic to the source of mAb2 (BALB/c).

Age-dependent penetrance of disease in a transgenic mouse model of familial amyotrophic lateral sclerosis.

The mutation gly93-->ala of Cu,Zn superoxide dismutase (SOD) is found in patients with familial amyotrophic lateral sclerosis and causes motor neuron disease when expressed in transgenic mice. The progression of clinical and pathological disease was studied in a line of mice designated G1H. Clinical disease started at 91 +/- 14 days of age with fine shaking of the limbs, followed by paralysis and death by 136 +/- 7 days of age. Pathological changes begin by 37 days of age with vacuoles derived from swollen mitochondria accumulating in motor neurons. At the onset of clinical disease (90 days), significant death of somatic motor neurons innervating limb muscles has occurred; mice at end-stage disease (136 days) show up to 50% loss of cervical and lumbar motor neurons. However, neither thoracic nor cranial motor neurons show appreciable loss despite vacuolar changes. Autonomic motor neurons also are not affected. Mice that express wild-type human Cu,Zn SOD remain free of disease, indicating that mutations cause neuron loss by a gain-of-function. Thus, the age-dependent penetrance of motor neuron disease in this transgenic model is due to the gradual accumulation of pathological damage in select populations of cholinergic neurons.

Immunohistochemical detection of neural cell adhesion molecule and laminin in X-linked dystrophic dogs and mdx mice.

Although dystrophin deficiency is known to be the genetic and biochemical defect causing Duchenne muscular dystrophy (DMD), much remains unknown about the underlying factors affecting clinical and pathological expression of the disease. Two animal forms of muscular dystrophy resembling DMD have been described. Neural cell adhesion molecule (NCAM) and laminin expression were examined in the proliferation-competent mdx mouse and non-regenerative "golden retriever muscular dystrophy dog" (GRMD). The results showed that (1) NCAM expression was greater in dystrophic dogs and mice than in age-matched normal animals, (2) myoblast-specific NCAM was greater in mdx mice than in dystrophic dogs, and (3) laminin strongly labelled mdx and GRMD myofibre membranes but was also sometimes found in individual interstitial cells of mdx muscle. Expression of these proteins may partly determine the clinicopathological expression of dystrophin deficiency.

Magnetic affinity cell sorting (MACS) separation and flow cytometric characterization of neural cell adhesion molecule-positive, cultured myogenic cells from normal and dystrophic dogs.

We developed a magnetic affinity cell sorting (MACS) assay based on differential expression of neural cell adhesion molecule (NCAM) isoforms in muscle cell cultures from normal and dystrophic dogs. NCAM is expressed during normal muscle differentiation, but has not been extensively examined within the context of muscle disease. A myogenic MACS assay could potentially maximize chances of obtaining normal nonsenescent, low-passage myogenic cells capable of proliferating in vitro and in vivo following transplantation. Myoblast-specific anti-NCAM polyclonal antibody directed against the NCAM isoform associated with muscle cell proliferation more effectively separated mixed canine cultures than did monoclonal antibodies directed against differentiated NCAM isoforms in the MACS assay. Flow cytometry using 5.1H11 anti-NCAM monoclonal antibody was then performed on normal and dystrophic fractionated cells and the results from these two groups were compared to each other and to nonfractionated cell populations. Normal canine cell cultures that had not been separated contained a larger percentage of FACscan-positive cells than did corresponding dystrophic canine cell cultures. Prior polyclonal anti-NCAM MACS separation of dystrophic cultures yielded higher numbers of adherent cells and higher gating percentages of 5.1H11-positive cell populations than did normal cultures. However, cells from dystrophic animals exhibited lower mean fluorescent expression of NCAM than normal cells.

A retrospective study of disease and mortality in zebra finches.

Few published reports exist describing morbidity and mortality in domestic zebra finch colonies maintained in a laboratory animal setting. A retrospective study of clinical disease and mortality in quarantined adult zebra finches was performed. Animals were observed during the 2 week quarantine period and for at least 1 month afterwards (42 days). Signs of disease, including feather and beak abnormalities, oculonasal discharge, increased respiratory rate or stridor, abdominal enlargement, pasty vent, diarrhea, lameness and pectoral muscle loss, were evaluated in our colony during this time. History, physical examination, laboratory testing and postmortem evaluation were used to determine causes of clinical disease. Common clinical findings in sick finches included sudden death, ruffled feathers, increased respiratory rate or gape mouthed breathing, pasty vent or frank diarrhea, and beak discoloration. Organisms frequently isolated were Staphylococcus spp., E. coli, Enterobacter spp., and Coccidia spp. Of the finches that died while in the colony (29.5%), 23.0% died in the first week after arrival. Pathogens frequently isolated from tissues cultured at necropsy included: E. coli, Staphylococcus aureus, Enterobacter spp., and Candida albicans. When observed, pathological lesions consisted of air sacculitis, fibrinopurulent polyserositis and ventriculitis.

Detection of mouse thymic virus (MTLV) antigens in infected thymus by competition immunoassay.

Mouse thymic virus (MTLV; murid herpesvirus 3) causes T lymphocyte depletion, thymic necrosis and immunosuppression in acutely infected neonatal mice. Infected animals shed virus persistently in saliva for prolonged periods of time. The standard procedure for detection of MTLV in infected mice is an in vivo infectivity assay. A sensitive enzyme linked immunosorbent assay (ELISA) recently has been developed for the detection of antibodies to MTLV. However, a direct test for viral antigen would be desirable in order to identify animals shedding virus, in the event that some infected animals may remain seronegative. For this purpose, we developed and assessed the feasibility of a competition ELISA to detect viral antigens in MTLV infected tissue. In competition ELISA, unknown tissue samples possibly containing antigen are mixed with a known quantity of antibody. Subsequent measurement of unbound antibody by ELISA is used to determine the presence of MTLV antigen in the samples. By competition ELISA we were able to detect MTLV in thymuses of acutely infected animals at a level of approximately 16 ID50 per sample. This technique shows promise as an alternative to infectivity testing methods and is also useful for confirmatory tests in MTLV serology.