Anti-arthropod saliva antibodies among residents of a community at high risk for Lyme disease in California.

The role of the western black-legged tick (Ixodes pacificus) versus that of other potential arthropod vectors in the epidemiology of Lyme disease was evaluated by determining the prevalence of anti-arthropod saliva antibodies (AASA) among residents (n = 104) of a community at high-risk (CHR). Salivary gland extracts prepared from I. pacificus, the Pacific Coast tick (Dermacentor occidentalis), the western cone-nose bug (Triatoma protracta), and the western tree-hole mosquito (Aedes sierrensis) were used as antigens in an ELISA. Sera from 50 residents of the San Francisco Bay region in northern California and 51 residents of Imperial County in southern California served as comparison groups. The prevalence of AASA ranged from 2% for A. sierrensis to 79% for I. pacificus in study subjects, 0% for D. occidentalis to 36% for I. pacificus among residents of the San Francisco Bay region, and 6% for I. pacificus to 24% for A. sierrensis in residents of Imperial County. The associations between AASA and demographic factors, potential risk factors, probable Lyme disease, and seropositivity for Borrelia burgdorferi were assessed for 85 members of the CHR. Seropositivity for I. pacificus and B. burgdorferi were significantly correlated, the relative risk of seropositivity to B. burgdorferi was about 5 (31% versus 6%) for subjects who were seroreactive to I. pacificus, nearly every individual who was seropositive for B. burgdorferi had elevated levels of antibodies to I. pacificus, and the mean titer for antibodies to I. pacificus was significantly higher for subjects seropositive versus those seronegative for B. burgdorferi. Together, these findings support the widely held belief that I. pacificus is the primary vector of B. burgdorferi for humans in northern California, and they demonstrate the utility of the AASA method as an epidemiologic tool for studying emerging tick-borne infections.

Characterization of an insulin degrading enzyme from cultured human lymphocytes.

An insulin degrading enzyme from cultured human lymphocytes, IM-9 cells, has been purified and characterized. The biochemical, enzymatic and immunological characteristics of this enzyme were all found to be similar to the characteristics of insulin degrading enzymes previously isolated from rat and pig skeletal muscle. Furthermore, this insulin degrading enzyme was found to have no effect on the structure of the insulin receptor nor to be linked to the insulin receptor either on the plasma membrane of cells or when they are shed into the media. The present studies suggest that the IM-9 lymphocytes, which have been extensively used to study the human insulin receptor, may also be a good system for studying human insulin degrading enzymes.

Production and characterization of a monoclonal antibody to rat liver thiol: protein-disulfide oxidoreductase/glutathione-insulin transhydrogenase.

Rat liver thiol:protein-disulfide oxidoreductase/glutathione-insulin transhydrogenase (glutathione:protein disulfide oxidoreductase, EC 1.8.4.2) was purified and found to give two bands on sodium dodecyl sulfate polyacrylamide gel electrophoresis. A monoclonal antibody was produced against this enzyme preparation and found to remove all the insulin degrading activity of purified preparations of the enzyme. This monoclonal antibody was also found to react with the two different forms of the enzyme observed on gel electrophoresis. These results suggest that glutathione-insulin transhydrogenase can exist in more than one state.

Degradation of insulin-like growth factors I and II by a human insulin degrading enzyme.

A human insulin degrading enzyme purified from IM-9 lymphocytes was tested for its ability to degrade insulin-like growth factor I (IGF-I) and insulin-like growth factor II (IGF-II). Degradation of these molecules was assessed by trichloroacetic acid precipitation, binding to specific receptors and chromatography on Sephadex G-50. All three techniques indicated that the enzyme readily degraded IGF-II and slightly degraded IGF-I. The IGF-II degrading activity chromatofocused with the insulin degrading activity and was absorbed by specific antibodies to the insulin degrading enzyme. These studies indicate, therefore, that a human insulin degrading enzyme can degrade IGF-II and, to a lesser extent, IGF-I.

Preferential degradation of the beta subunit of purified insulin receptor. Effect on insulin binding and protein kinase activities of the receptor.

Collagenase preparations (a mixture of enzymes including collagenase, clostripain, and a casein-degrading protease) degraded the beta subunit (Mr = 95,000) of the purified insulin receptor into fragments of Mr less than 15,000, without degrading the alpha subunit. The resulting beta-digested insulin receptor preparations were found to bind insulin as well as control insulin receptor, as assessed by either cross-linking of 125I-insulin to the digested receptor or by separating insulin bound to receptor from free insulin by high performance liquid chromatography. Moreover, the beta-digested insulin receptor preparations were still precipitated by a monoclonal antibody directed against the insulin-binding site. In contrast, the beta-digested insulin receptor lacked protein kinase activity since it no longer phosphorylated either itself, or an exogenous substrate, calf thymus histone. These results support the identification of the beta subunit of the insulin receptor as a protein kinase.