Family studies of association between HLA and specific immune responses to highly purified pollen allergens

In 76 members of 13 large families, we investigated whether association exists between specific familialHLA haplotypes and immune responsiveness to four different highly purified pollen antigens (ragweed antigens E, Ra3, and Ra5 and rye grass Group I). Specific immune response was studied quantitatively by measurement of IgE-mediated skin sensitivity, serum IgG antibody, and antigen-induced lymphocyte proliferationin vitro. We found no evidence for association between specificHLA haplotype and specific response measured by any one or more of our indices of immune function. In several families we found evidence of specific response in one generation but not in another. We found a number of instances of individuals exhibiting lymphocyte responsiveness to an antigen, but no detectable specific IgE or IgG antibody. Surprisingly, we also found a few cases of individuals with marked IgE and/or IgG responses to a given antigen who showed no measurable lymphocyte responsiveness to that antigen, despite lymphocyte responsiveness to other nonimmunologically crossreacting antigens. In several cases, we also observed lymphocyte stimulation and serum IgG antibody, but no detectable IgE response. Our results conflict with previous investigators' reports of linkage betweenHLA haplotype and specific IgE-mediated skin sensitivity in families.HLA- linked immune response (Ir) loci may exist in humans, but genetic complexity and the limits of current technology preclude our ability to demonstrate their existence.     

Weight and mortality in the Whitehall Study.

Ten-year mortality rates in men aged 40-64 years in the Whitehall Study were analysed in relation to weight and height at the initial examination. At ages 40-49 "all-causes" mortality increased with increasing body mass index; but this simple relation disappeared at older ages, where there was an increased mortality in the lowest quintile of body mass index. The "all-ages" relation was "J"-shaped, and this could not be explained by the confounding effects of blood pressure, cholesterol values, and cigarette smoking. Some, but not all, of the J shape was due to a high short-term mortality in thin men from cancers (presumably already present at examination). At younger ages mortality from coronary heart disease was positively related to body mass index, but this depended on its association with other risk factors. Mortality from causes other than cancers or coronary heart disease was highest in the lowest quintile of body mass index.     

Myocardial ischemia/reperfusion injury in the inducible nitric oxide synthase knockout mice.

Inducible nitric oxide synthase (iNOS) plays an important role in the inflammatory process of certain major cardiac disorders including myocardial infarction and allograft rejection. However, the role of iNOS in acute myocardial ischemia has not been well defined. We determined the effects of genetically disruption of the intact iNOS system on cardiac tolerance to ischemia/reperfusion injury. Adult male wild-type (WT) and iNOS knockout (KO) B6,129 mice were subjected to 20 min global ischemia and 30 min reperfusion in a Langendorff isolated perfused heart model (37 degrees C, n = 10/each group). Ventricular contractile function, heart rate, coronary flow, and leakage of intracellular enzymes (CK and LDH) were not significantly different between the groups during pre-ischemia as well as reperfusion period (P > 0.05). Myocardial infarct size was also not significantly different between WT (20.2+/-2.0% of risk area) and KO mice (23.5+/-3.8%; Mean+/-SEM, P > 0.05). However, the post-ischemic heart rate was significantly preserved in KO as compared to WT (P < 0.05). We conclude that disruption of iNOS gene does not exacerbate ischemia/ reperfusion injury in the heart.     

Phosphorylation of dystrophin and alpha-syntrophin by Ca(2+)-calmodulin dependent protein kinase II.

A Ca(2+)-calmodulin dependent protein kinase activity (DGC-PK) was previously shown to associate with skeletal muscle dystrophin glycoprotein complex (DGC) preparations, and phosphorylate dystrophin and a protein with the same electrophoretic mobility as alpha-syntrophin (R. Madhavan, H.W. Jarrett, Biochemistry 33 (1994) 5797-5804). Here, we show that DGC-PK and Ca(2+)-calmodulin dependent protein kinase II (CaM kinase II) phosphorylate a common site (RSDS(3616)) within the dystrophin C terminal domain that fits the consensus CaM kinase II phosphorylation motif (R/KXXS/T). Furthermore, both kinase activities phosphorylate exactly the same three fusion proteins (dystrophin fusions DysS7 and DysS9, and the syntrophin fusion) out of a panel of eight fusion proteins (representing nearly 100% of syntrophin and 80% of dystrophin protein sequences), demonstrating that DGC-PK and CaM kinase II have the same substrate specificity. Complementing these results, anti-CaM kinase II antibodies specifically stained purified DGC immobilized on nitrocellulose membranes. Renaturation of electrophoretically resolved DGC proteins revealed a single protein kinase band (M(r) approximately 60,000) that, like CaM kinase II, underwent Ca(2+)-calmodulin dependent autophosphorylation. Based on these observations, we conclude DGC-PK represents a dystrophin-/syntrophin-phosphorylating skeletal muscle isoform of CaM kinase II. We also show that phosphorylation of the dystrophin C terminal domain sequences inhibits their syntrophin binding in vitro, suggesting a regulatory role for phosphorylation.     

Feline leukemia virus DNA vaccine efficacy is enhanced by coadministration with interleukin-12 (IL-12) and IL-18 expression vectors

The expectation that cell-mediated immunity is important in the control of feline leukemia virus (FeLV) infection led us to test a DNA vaccine administered alone or with cytokines that favored the development of a Th1 immune response. The vaccine consisted of two plasmids, one expressing the gag/pol genes and the other expressing the env gene of FeLV-A/Glasgow-1. The genetic adjuvants were plasmids encoding the feline cytokines interleukin-12 (IL-12), IL-18, or gamma interferon (IFN-gamma). Kittens were immunized by three intramuscular inoculations of the FeLV DNA vaccine alone or in combination with plasmids expressing IFN-gamma, IL-12, or both IL-12 and IL-18. Control kittens were inoculated with empty plasmid. Following immunization, anti-FeLV antibodies were not detected in any kitten. Three weeks after the final immunization, the kittens were challenged by the intraperitoneal inoculation of FeLV-A/Glasgow-1 and were then monitored for a further 15 weeks for the presence of virus in plasma and, at the end of the trial, for latent virus in bone marrow. The vaccine consisting of FeLV DNA with the IL-12 and IL-18 genes conferred significant immunity, protecting completely against transient and persistent viremia, and in five of six kittens protecting against latent infection. None of the other vaccines provided significant protection.     

Segmental identity and cerebellar granule cell induction in rhombomere 1

Background  

Cerebellar granule cell precursors are specifically generated within the hindbrain segment, rhombomere 1, which is bounded rostrally by the midbrain/hindbrain isthmus and caudally by the boundary of the Hoxa2 expression domain. While graded signals from the isthmus have a demonstrable patterning role within this region, the significance of segmental identity for neuronal specification within rhombomere 1 is unexplored. We examined the response of granule cell precursors to the overexpression of Hoxa2, which normally determines patterns of development specific to the hindbrain. How much does the development of the cerebellum, a midbrain/hindbrain structure, reflect its neuromeric origin as a hindbrain segment?     

Longitudinal analysis of feline leukemia virus-specific cytotoxic T lymphocytes: correlation with recovery from infection

Feline leukemia virus (FeLV) is a common naturally occurring gammaretrovirus of domestic cats that is associated with degenerative diseases of the hematopoietic system, immunodeficiency, and neoplasia. Although the majority of cats exposed to FeLV develop a transient infection and recover, a proportion of cats become persistently viremic and many subsequently develop fatal diseases. To define the dominant host immune effector mechanisms responsible for the outcome of infection, we studied the longitudinal changes in FeLV-specific cytotoxic T lymphocytes (CTLs) in a group of na?ve cats following oronasal exposure to FeLV. Using (51)Cr release assays to measure ex vivo virus-specific cytotoxicity, the emerging virus-specific CTL response was correlated with modulations in viral burden as assessed by detection of infectious virus, FeLV p27 capsid antigen, and proviral DNA in the blood. High levels of circulating FeLV-specific effector CTLs appeared before virus neutralizing antibodies in cats that recovered from exposure to FeLV. In contrast, persistent viremia was associated with a silencing of virus-specific humoral and cell-mediated host immune effector mechanisms. A single transfer of between 2 x 10(7) and 1 x 10(8) autologous, antigen-activated lymphoblasts was associated with a downmodulation in viral burden in vivo. The results suggest an important role for FeLV-specific CTLs in retroviral immunity and demonstrate the potential to modulate disease outcome by the adoptive transfer of antigen-specific T cells in vivo.     

Short and long term consequences of increases in exotic species richness on water filtration by marine invertebrates

Although recent research has considered the consequences of global declines in the number of species, less attention has focused on the aggregate effects of regional increases in species richness as a result of human-mediated introductions. Here we examine several potential ecosystem consequences of increasing exotic species diversity of suspension feeding marine invertebrates. First, we experimentally manipulated native and non-native suspension feeder richness and measured its effect on short-term phytoplankton clearance rates. Multispecies communities all performed similarly, regardless of whether they were dominated by natives, exotics, or an even mix of the two. Individual species varied considerably in filtration rates, but non-native species often filtered less than the most similar native. Second, we determined potential changes in integrated function over time by comparing seasonal patterns of recruitment as a proxy for the ability to quickly recover filtration capacity after a disturbance. We found that exotic species have complementary seasonal phenologies both to native species and each other. Our results suggest that the consequences of local increases in species richness due to invasions may be manifest over long (annual to interannual) time scales, even when short term changes in ecosystem function are negligible.