Biomedical subjects

W C Lai

Publications and source records attributed to W C Lai.

31 records · Page 2Linked to original sources

PubMed1991

The outer membrane of Pasteurella multocida 3:A protects rabbits against homologous challenge.

The protective efficacy of a vaccine purified from the Pasteurella multocida 3:A outer membrane (OM) was evaluated in rabbits by homologous challenge. Twenty-seven rabbits were divided into four groups: 1, vaccinated with OM and challenged; 2, nonvaccinated and challenged; 3, vaccinated with OM only; and 4, nonvaccinated and not challenged. Rabbits were immunized intranasally with 1 mg of OM protein on days 0, 7, 14, and 35, challenged intranasally on day 49, and killed on day 63. Mortality rates were 0, 67, 0, and 0% for groups 1 through 4, respectively. The prevalence of pneumonia was reduced from 73 (group 2) to 20% (group 1). The severity of pneumonia was reduced from 0.62 (group 2) to 0.07 (group 1), as measured by the group lesion index. The number of P. multocida in nasal cavities was reduced from 3.89 x 10(5) (group 2) to 6.19 x 10(2) (group 1). The geometric mean number of P. multocida in lungs was 8,360,000-fold less in group 1 than in group 2. Similarly, the prevalence of P. multocida colonization in nonrespiratory organs was reduced from 47 (group 2) to 4% (group 1). Furthermore, group 1 and 3 rabbits developed significantly elevated immunoglobulin A antibodies in nasal secretions and lung lavages and significantly elevated immunoglobulin G antibodies in lung lavages and sera. In addition, rabbit immune sera contained antibodies against P. multocida OM proteins and lipopolysaccharides and inhibited P. multocida proliferation in mouse lungs. These results indicate that a vaccine prepared from the OM of P. multocida provides a significant protection in rabbits against homologous challenge.

Animals↗

PubMed1991

Antibodies to outer membrane proteins but not to lipopolysaccharide inhibit pulmonary proliferation of Pasteurella multocida in mice.

The role of rabbit antibodies against Pasteurella multocida outer membrane proteins and lipopolysaccharides (LPS) in resistance remains unknown. Pooled immune sera against P. multocida outer membranes were prepared from specific-pathogen-free rabbits immunized with sucrose gradient-purified P. multocida outer membranes. Western immunoblotting showed that purified outer membrane protein antibodies reacted strongly against the outer membrane proteins but not the purified LPS. Affinity-purified LPS antibodies exhibited strong reactivity against purified LPS and very little reactivity against outer membrane vesicles. Mice were inoculated intranasally with immune serum or normal rabbit serum, challenged intranasally with 10(6) CFU of P. multocida, and euthanatized 48 h later to determine the number of P. multocida organisms in the lungs. Mice inoculated with pooled immune serum had a 3,300-fold reduction (P less than 0.001) in the numbers of P. multocida in the lungs as compared with the controls. Similarly, mice inoculated with purified outer membrane protein antibodies had a 201-fold reduction (P less than 0.001) in the numbers of P. multocida. Conversely, mice inoculated with affinity-purified LPS antibodies had a 1.1-fold reduction (P greater than 0.50) in the numbers of P. multocida. These results show that antibodies against the outer membrane proteins but not the LPS are the components of rabbit immune sera which inhibit P. multocida proliferation in mouse lungs.

Animals↗

PubMed1991

Protease-like sequence in hepatitis B virus core antigen is not involved in the cleavage processes of core protein in Escherichia coli.

A DNA fragment, coding for hepatitis core antigen (HBcAg), was amplified by polymerase chain reaction and inserted into a lambda PL promoter-derived expression vector. The recombinant plasmid was transformed into Escherichia coli and proteins produced after heat induction were analyzed. In addition to the 21 kDa HBcAg protein, several smaller related polypeptides, particularly one of 17 kDa in size, were also detected with rabbit anti-HBcAg antiserum. Whether the protease-like sequence of core protein involved in the self-cleavage process to form the 17 kDa polypeptide was investigated by a deletion experiment. Our results with a mutant in which 7 amino acids of the conserved protease-like region in the core protein have been deleted suggest that the cleavage does not depend on the presence of these protease-like sequence. In addition, the core protein synthesized from in vitro translation reaction was not cleaved. Core particles from E. coli lysate were purified by sucrose and cesium chloride density gradient centrifugations and subsequently treated with 0.2% of SDS and 0.2% of beta-mecaptoethanol. Immunoblotting analysis, however, did not reveal any conversion of the 21 kDa protein to smaller ones. In conclusion, our results suggest that the protease-like domain at the N-terminus of the core protein does not contain intrinsic autocleavage activity, nor could the HBcAg be converted to smaller antigens by detergent treatment.

Aspartic Acid Endopeptidases↗

PubMed1990

Resistance to Mycoplasma pulmonis mediated by activated natural killer cells.

Infection of C57BL/6J mice with Mycoplasma pulmonis (MP) enhanced NK cell function 3-7 days later, as detected by in vitro and in vivo assays. Moreover, spleen and lung cells of acutely infected C57BL/6J mice inhibited MP growth in vitro. The effectors were eliminated by treatment with anti-NK antibody in vivo and anti-asialo GM1 serum or anti-3A4 antibody plus complement in vitro. Clearance of viable and radiolabeled MP from the lungs was also enhanced in acutely infected mice. Acutely infected mice with severe combined immunodeficiency (SCID) eliminated viable MP faster than did uninfected mice. Antibodies to interferon-gamma (IFN-gamma) impaired clearance of MP from the lungs of SCID mice and decreased their survival times. Activated NK cells can function in resistance to early stages of infection with MP. NK cells directly inhibit MP with secrete IFN-gamma, which may activate macrophages or inhibit the growth of MP or both.

Animals↗

PubMed1990

Biological evaluation of Mycoplasma pulmonis temperature-sensitive mutants for use as possible rodent vaccines.

Temperature-sensitive mutants (TSMs) of Mycoplasma pulmonis were produced by treating the wild-type strain with N-methyl-N'-nitro-N-nitrosoguanidine. Three TSMs were selected at 38 degrees C, as a restrictive temperature, and at 34 degrees C, as a permissive temperature. Two TSMs, UTCMI and UTCMII, were proven to be nonpathogenic but immunogenic. In addition, they did not induce pneumonia, tracheitis, or tympanitis but did induce mild rhinitis. They were stable after 10 passages in vitro and in vivo. They elicited excellent antibody production and cell-mediated immunity in vaccinated rats. They also were not mitogenic to rat lymphocytes. Rats immunized intranasally with these TSMs were significantly protected against challenge with wild-type organisms. These mutants were morphologically and serologically indistinguishable from the wild-type organisms. The growth characteristics and antibiotic sensitivities were similar to those of wild-type organisms, except that they grew only at 34 degrees C. In contrast to wild-type organisms, they did not bind to or lyse sheep erythrocytes. Thus, these TSMs may qualify as a vaccine to prevent M. pulmonis infection in rats.

Animals↗

PubMed1989

Mycoplasma pulmonis depresses humoral and cell-mediated responses in mice.

Humoral and cell-mediated immune responses to sheep red blood cells (SRBC) were studied in mice infected experimentally with Mycoplasma pulmonis. The hemagglutinating (HA) antibody against SRBC was evaluated at 0, 3, 5, 7, 14, 21 and 28 days postinfection (PI). Antibody tiers during all days PI were depressed significantly (p less than 0.05) in infected mice as compared to noninfected controls. The HA antibody, which is of the IgM class, peaks at day 5 PI. There is no shift in the kinetics of the humoral response in M. pulmonis infected mice. Cellular immune responses were evaluated by a delayed-type hypersensitivity (DTH) reaction and the lymphocyte transformation technique. Mice were sensitized at 0,3,5,7,14, 21 and 28 days PI with SRBC and challenged by footpad injection of SRBC 7 days later. The DTH reaction measured at 24 hours after challenge was depressed significantly (p less than 0.05) in all infected animals. After a transient enhancement on day 3 PI, the DTH responses remained depressed through day 28 PI. The lymphocyte transformation test showed a significantly (p less than 0.05) depressed response except on days 5 and 7 PI. These results indicate that M. pulmonis infection in mice suppresses the humoral antibody and cell-mediated immune responses.

Animals↗

PubMed1988

Experimental Mycoplasma pulmonis infection of rats suppresses humoral but not cellular immune response.

Humoral antibody response to sheep red blood cells and cellular immune response to bovine serum albumin were studied in Mycoplasma pulmonis infected, adult, male Sprague-Dawley rats. The hemagglutinating antibody response to sheep red blood cells was evaluated at 0, 3, 5, 7, 14, 21 and 28 days postinfection. Antibody titers during all days postinfection were depressed significantly (p less than 0.05) in infected rats as compared to noninfected controls. Cellular immune responses were evaluated by a delayed hypersensitivity response. Rats were sensitized at 0, 3, 5, 7, 14, 21 or 28 days postinfection with bovine serum albumin and challenged with heat aggregated bovine serum albumin 7 days later. Cell-mediated immune responses in infected rats were not significantly different at any point from controls. These results indicate that M. pulmonis infection in rats suppresses the humoral antibody response to sheep red blood cells, but not the cellular immune response to bovine serum albumin.

Animals↗

PubMed1988

The effect of transportation stress on splenic natural killer cell activity in C57BL/6J mice.

Splenic natural killer cell activity and plasma corticosterone levels were measured in air- and truck-transported C57BL/6J mice (Mus musculus) on days 0, 1, 3 and 5 post-arrival. These data are important in determining adequate stabilization periods for transported animals before studies involving natural killer cells are begun. Three control groups (phosphate buffered saline, polyinosinic-polycytidylic acid, and hydrocortisone injected mice) were stabilized in the animal facilities 3 weeks before the start of experiments. Natural killer activity in transported mice was reduced significantly (p less than 0.05) on day 0 and returned to normal levels by 24 hours. Plasma corticosterone levels were increased significantly (p less than 0.005) on day 0 and returned to control levels by day 1, correlating inversely with splenic natural killer activity. This study indicates that stress resulting from transportation causes a short-term decrease in the splenic natural killer cell activity of mice, and this decrease may be related to the increased plasma corticosterone levels induced by the stressful event. We conclude that mice should be stabilized at least 24 hours before experiments involving the natural killer cell system are begun.

Animals↗

PubMed1987

Mycoplasma pulmonis infection augments natural killer cell activity in mice.

The goal of this study was to determine if experimental Mycoplasma pulmonis infection augmented splenic natural killer (NK) cell activity in mice. A 4 hour 51Cr-release in vitro assay using YAC-1 tumor target cells was employed to measure splenic NK cell activity in C57BL/6J mice infected intraperitoneally with M. pulmonis and in uninfected controls. Transient augmentation of the NK cells was observed, peaking at day 3 postinoculation (PI) and gradually returning to normal levels by day 10 PI. Selective depletion studies showed that the cells responsible for killing target cells were NK cells. They were nonadherent to nylon wool, not susceptible to Thy-1.2 antibody and susceptible to asialo GM1 ganglioside antibody. Inadvertent augmentation of the NK cell system due to M. pulmonis infection may complicate the interpretation of research data, especially in immunology and cancer studies.

Animals↗

PubMed1986

Comparison of Chalquest and Hayflick media, with and without ammonium reineckate, for isolating Mycoplasma pulmonis from rats.

Chalquest and Hayflick media with and without ammonium reineckate were compared for isolation of Mycoplasma pulmonis from the nasopharyngeal ducts, tracheobronchial trees, and middle ears of 66 naturally infected rats. The results show that 92% (366 of 396) of the samples were positive for M. pulmonis in Chalquest medium with and without ammonium reineckate and 66% (260 of 396) were positive in Hayflick medium with and without ammonium reineckate (P less than 0.001). An enhancing effect of ammonium reineckate on M. pulmonis isolation was observed only in Hayflick medium; the isolation rate was 76% (151 of 198) in Hayflick medium with ammonium reineckate as compared with 55% (109 of 198) in Hayflick medium without ammonium reineckate (P less than 0.03). No significant differences in isolation rates were observed between Chalquest medium with and without ammonium reineckate. The mean growth time of M. pulmonis on Chalquest medium was 3.4 days as compared with 5.1 days in Hayflick medium, indicating that M. pulmonis can be detected earlier on Chalquest medium than on Hayflick medium. These data indicate that Chalquest medium is superior to Hayflick medium for M. pulmonis isolation from rats.

Animals↗

PubMed1985

Carbon immunoassay--a simple and rapid serodiagnostic test for feline toxoplasmosis.

A serodiagnostic test, simpler and more rapid to perform than traditional methods, was sought to identify Toxoplasma gondii antibody in research cats. The reliability and sensitivity of the direct and indirect carbon immunoassay (CIA) tests were compared to each other and to the indirect immunofluorescent antibody (IFA) test. The three tests were used to detect the presence or absence of T. gondii antibody in the sera of 94 cats. The results of this study show that the CIA tests correlate with one another and the IFA test nearly 99%, indicating they are highly reliable. Comparison of titers of the positive sera indicate a high degree of sensitivity as well.

Animals↗

PubMedDate not supplied

Molecular characterization of the rat NK cell receptor 2B4.

2B4 (CD244) is a cell surface glycoprotein of the immunoglobulin superfamily involved in the regulation of natural killer and T lymphocyte function. It is the high affinity counter-receptor for CD48. In mouse and human NK cells, crosslinking of 2B4 with a specific monoclonal antibody or with CD48 can trigger cell-mediated cytotoxicity, IFN-gamma secretion, phosphoinositol turnover and NK cell invasiveness. Recent reports of defective 2B4 signaling and NK cell function in X-linked lymphoproliferative syndrome suggest that this may contribute to the progression of this human disease. Here we describe the molecular characterization of the rat 2B4 gene. The cDNA encodes a protein of 395 amino acid residues that contain two Ig domains in the extracellular region and three unique tyrosine motifs (TxYxxV/I/A) in the cytoplasmic region. The predicted protein has 81 and 68% similarity with mouse 2B4 and human 2B4, respectively. Additionally, it has 94 and 89% similarity at the protein level with the recently reported rat 2B4 related genes, r2B4R-tm and r2B4R-se respectively. Northern blot analysis indicated the presence of multiple transcripts in rat LAK cells and RNK-16 cells. Immunoprecipitation and deglycosylation studies showed that rat 2B4 is glycosylated to similar extent as that of mouse and human 2B4. The cloning of r2B4 in the light of the availability of rat NK cell lines should facilitate in vitro and in vivo experiments to decipher the functional role of 2B4 in NK cell biology.

Amino Acid Sequence↗

PubMedDate not supplied

Natural resistance to Mycoplasma pulmonis infection in mice: host resistance gene(s) map to chromosome 4.

Strains of mice differ greatly in resistance to infection in their lungs with virulent Mycoplasma pulmonis (MP) organisms even during the first 5 days, prior to detection of humoral or T cell mediated acquired immune responses. C57BL/6 mice are resistant, and BALB/c and C3H mice are susceptible, and one major gene, MP, not linked to the H2 major histocompatibility complex, regulates resistance. C57BL/6 x C3H (B x H) and BALB/c x C57BL/6 (C x B) recombinant inbred strain mice were infected intratracheally with the T2 strain of MP. Five days later, the recovery of organisms from tracheolung lavages and lung tissue was determined. The strain distribution pattern of resistance indicated that the MP gene maps to chromosome 4. B6.C-H18 (B6 mice congenic for the BALB/c H18 gene of chromosome 4) were much more susceptible than B6 mice, but were less susceptible than BALB/c mice, supporting the data obtained with the recombinant inbred strain mice, but suggesting that other genes may also influence resistance to infection with MP.

Animals↗