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P Mottram

Publications and source records attributed to P Mottram.

17 recordsLinked to original sources

Anti-Gal antibody-mediated allograft rejection in alpha1,3-galactosyltransferase gene knockout mice: a model of delayed xenograft rejection.

BACKGROUND: The key role of anti-galactose alpha1,3-galactose (anti-alphaGal) xenoantibodies in initiating hyperacute xenograft rejection has been clearly demonstrated using a variety of in vitro and in vivo approaches. However, the role of anti-alphaGal antibodies in mediating post-hyperacute rejection mechanisms, such as antibody-dependent cellular cytoxicity, remains to be determined, primarily because of the lack of a small animal model with which to study this phenomena. METHODS: Hearts from wild-type mice were transplanted heterotopically into alpha1,3-galactosyltransferase knockout (Gal KO) mice, which like humans develop antibodies to the disaccharide galactose alpha1,3-galactose (Gal). At the time of rejection, hearts were examined histologically to determine the mechanism of rejection. RESULTS: Hearts from wild-type mice transplanted into high-titer anti-alphaGal recipients were rejected in 8-13 days. Histological examination demonstrated a cellular infiltrate consisting of macrophages (80-90%), natural killer cells (5-10%), and T cells (1-5%). In contrast, wild-type hearts transplanted into low anti-Gal titer recipients demonstrated prolonged (>90 day) survival. However, a significant proportion (30-40%) of these underwent a minor rejection episode between 10 and 13 days, but then recovered ("accommodated"). CONCLUSIONS: The results of this study suggest that the Gal KO mouse is a useful small animal vascularized allograft model, in which the role of anti-alphaGal antibody in graft rejection can be studied in isolation from other rejection mechanisms. The titer of anti-alphaGal antibody was found to be the critical determinant of rejection. The histopathological features of rejection in this model are very similar to other models of delayed xenograft rejection, in both the timing and composition of the cellular infiltrate. The Gal KO mouse therefore provides a new rodent model, which will aid in the identification of the distinct components involved in the pathogenesis of delayed xenograft rejection.

Animals

Effect of IL-4 deletion on cardiac allograft survival in the BALB/c to 129Sv x C57BL/6 strain combination.

To investigate the effect of IL-4 deletion on cardiac allograft survival, vascularized BALB/c cardiac allografts were placed in C57BL/6, 129Sv x C57BL/6 (IL-4 +/+) or 129Sv x C57BL/6 IL-4 knockout mice (IL-4-/-). Untreated recipients rejected allografts in < 15 days while isografts survived indefinitely (> 100 days). Treatment with anti-CD4 (GK1.5) for 4 days at the time of allografting increased mean survival to > 100 days in C57BL/6, 90+/-16 days in 129Sv x C57BL/6 (IL-4 +/+) and 68 +/- 36 days in 129Sv x C57BL/6 (IL-4-/-) recipients. Although there was a trend towards shorter survival times in the IL-4-/- mAb-treated mice, survival in the three recipient groups was not significantly different (P = 0.07). A 30-day course of anti-CD4 did not further prolong BALB/c heart survival. All long-surviving hearts had histological evidence of parenchymal damage and transplant vascular disease. None of these recipients developed antigen-specific tolerance, since both donor and third party skin graft challenges were rejected when challenged at > 60 days post-graft and all primary grafts failed by 120 days. Thus the effects of IL-4 deletion were subtle and were seen only with low doses of immunosuppression in this high responder strain combination.

Animals

Development and survival of immature Culex annulirostris mosquitoes in southeast Queensland.

The rates of development and survival of the immature stages of Culex annulirostris were studied in three different breeding sites in the Brisbane area of southeast Queensland: a temporary rain-filled pool (TP), a semi-permanent pool (SPP), and an area of flooded grassland (FG) arising from the overflow of a permanent pond. Parallel observations were made on larvae and pupae either exposed to predation or in predator-free cages of 12 and 15 consecutive days respectively. Development was fastest and survival highest in the warmer TP. Average daily mortality of larvae (L) and pupae (P) was TP: L 8.3%, P 9.5%; SPP: L 25.2%, P 34.6%; FG: L 18.8%, P 35.4%. From life-tables and survivorship curves, survival from first instar to adult was TP 34.8%, SPP 2.0% and FG 4.2%. Mortality from predation was TP 43.2%, SPP 68.7% and FG 69.1%. Predator density was TP 0.32, SPP 1.48 and FG 1.76 per 300 ml sample.

Animals

Larval diet and the vector competence of Culex annulirostris (Diptera: Culicidae) for Murray Valley encephalitis virus.

Culex annulirostris Skuse larvae were reared on two different amounts of powdered dog chow and yeast, low intake (2.8 micrograms/larva) and high intake (8.4 micrograms/larva), to produce adults with mean wing lengths of 3.25 and 3.7 mm, respectively. The resulting adults were fed different dosages of Murray Valley encephalitis virus and after 10 d extrinsic incubation at 28 degrees C, the infection rate, transmission rate, salivary gland, and body titers were determined. There were no significant differences in any of these parameters.

Animals

Rearing temperature influences flavivirus vector competence of mosquitoes.

Culex annulirostris Skuse mosquitoes (Brisbane strain) were reared at 20 degrees C or 27 degrees C and the adult females were experimentally infected by feeding Murray Valley encephalitis virus (MVE). They were then maintained (a) in the insectary at 20 degrees C, after rearing at either 20 degrees C or 27 degrees C; (b) at ambient outdoor temperatures, range 12.2-28.9 degrees C, mean 19.6 degrees C; or (c) at 27 degrees C after rearing at 27 degrees C. There was no significant difference in rates of MVE infection or transmission when mosquitoes were reared and maintained constantly at 20 degrees C or 27 degrees C. However, for females kept at reduced temperature (i.e. mean = 19.6 degrees C or 20 degrees C after rearing at 27 degrees C), the infection and transmission rates of MVE were significantly reduced (2 x 8 replicates). This investigation illustrates that vector competence is depressed by decreasing temperatures for adult mosquitoes compared with those they experienced during development. Similar patterns were evident with previously published work on Japanese and St Louis encephalitis, dengue and yellow fever. The process appears to be reversible, i.e. increased temperature raises virus infection and transmission rates. It is concluded that, without incubation at warmer temperatures, flavivirus recovery from overwintering mosquitoes will be negatively biased.

Animals

The vector competence of Culex annulirostris, Aedes sagax and Aedes alboannulatus for Murray Valley encephalitis virus at different temperatures.

Culex annulirostris Skuse, colonized from Brisbane, Queensland, and Mildura, Victoria, Australia, were effective vectors of Murray Valley encephalitis virus at 20, 27 and 32-35 degrees C with full extrinsic incubation periods of 15, 10 and 4 days respectively. At 20 degrees C, 7-11 days post-infection, transmission by the Mildura colony (0-20%) was less efficient than the Brisbane colony (30-70%) but both were capable of 75-100% transmission after longer extrinsic incubation periods. Discriminant analysis of body and salivary gland titres showed that these were not satisfactory indicators of transmission. Wild-caught Aedes sagax (Skuse) and Cx annulirostris from the Murray Valley showed equal competence, but Aedes alboannulatus (Macquart) was a poor vector. The results provide data on rural amplification of Murray Valley encephalitis virus during spring and suggest that further work on the potential of Ae. sagax as a natural vector is warranted.

Aedes

In-vivo staining of Aedes vigilax, Aedes aegypti and Culex annulirostris larvae with Giemsa and other vital dyes.

Of 11 stains tested in the laboratory with larval Aedes aegypti, Giemsa at dosages of 6 mg/liter for 24 hr and 21 mg/liter for 3 hr proved most satisfactory for in-vivo staining. At these rates, Giemsa caused some mortality especially in first instars of Ae. vigilax, Ae. aegypti and Culex annulirostris but effectively was retained in all adults examined up to 3 days posteclosion. The method was deemed satisfactory for investigations of larval movement of Ae. vigilax, Cx. sitiens and Ae. alternans in salt marsh in southeast Queensland.

Aedes

Autogeny in Culex annulirostris from Australia.

In the laboratory, 8.5% and 5.1% of colonized Culex annulirostris from Brisbane and Mildura, Australia respectively were autogenous when reared and maintained on nutrient rich diets. Females reared and/or maintained on poor diets mainly had ovaries at Christophers' stage I and exhibited from 0 to 0.7% autogeny. All autogenous females had previously mated. Insemination rates in the Brisbane and Mildura colonies respectively, were 72.8% and 78.8%. No autogeny was detected in 997 females reared from 7 localities throughout Queensland but this may have been due to their poorer nutritional status (as indicated by wing size) or more likely to a low insemination rate of 0 to 16%. Our laboratory results, particularly with well-fed females, may have little relevance to the field situation, where adults are generally smaller and less well-nourished.

Animals

Influence of thymus genotype on acquisition of responsiveness in delayed-type hypersensitivity.

Antigen-pulsed macrophages (Mph) could sensitize syngeneic mice for delayed-type hypersensitivity (DTH) and also elicit sensitivity from mice sensitized to antigen in adjuvant provided these were syngeneic or semi-allogeneic to the strain providing the Mph. Sensitivity could not be elicited with antigen-pulsed allogeneic Mph. Antigen-pulsed Mph from low-responder (LR) strains could not sensitize LR mice nor F1 hybrids between responder (R) and LR strains. Normal F1 mice could be sensitized to respond to antigen presented on Mph or either parental type (i.e. P1 or P2): if, however, they were sensitized to antigen on P1 Mph, DTH transfer was restricted to naive P1 mice, not to P2 (restriction imposed by priming). F1 T cells derived from stem cells differentiating in a P1 thymus graft could be sensitized but could transfer sensitivity only to naive P1 mice, not to P2 (restriction imposed in thymus). When an antigen under Ir gene control was used, LR derived T cells differentiating in an (R X LR)F1 thymus could be sensitized but only if antigen was presented on (R X LR)F1 Mph not on LR Mph. Totally allogeneic chimaeras could be sensitized but only if given antigen in association with the appropriate Mph. These findings suggest that restriction of T cell activities can be imposed as a result of priming in some cases and as a result of differentiation within the thymus in others. LR strains appear to have a lesion at the level of antigen presentation by Mph; whether they also have a defect at the level of generation of T cell repertoire cannot be determined from the present investigations.

Animals

Major histocompatibility complex gene products on macrophages influence T cell activation.

Antigen-pulsed macrophages were used to sensitize or elicit sensitivity from mice of different strains to a variety of antigens. The results indicate that sensitization is directed, not to antigen as such, but to a complex structure on the macrophage surface determined partly by the antigen, and partly by a product coded by the major histocompatibility complex. Delayed type hypersensitivity could be provoked by antigen in responder (R) mice and in the F1 between responder and low responder (LR) strains, but not in LR mice unless pretreated by cyclophosphamide. Sensitivity could be transferred to naive LR-strain mice by lymph node cells taken 5 days after sensitization of cyclophosphamide-pretreated LR mice but not of F1 hybrids between LR and R strains. Sensitivity from these could be transferred only to naive F1 or R-strain mice. The results suggest that low responsiveness cannot be accounted for solely in terms of the operation of a cyclophosphamide-sensitive suppressor mechanism. It is postulated that antigen is less immunogenic when presented by LR-strain cells than by R-strain cells.

Animals

Declining blues.

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Age Factors

Distress signals.

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Age Factors