Identification of monoclonal antibodies that distinguish between 17D-204 and other strains of yellow fever virus.

Eight monoclonal antibodies (MAbs) prepared against the flaviviruses Saint Louis encephalitis, dengue 2 and dengue 3 viruses all recognized epitopes on the envelope protein of the prototype flavivirus, yellow fever (YF) virus. Three of these MAbs with flavivirus group-common specificity and two MAbs with a flavivirus-subgroup specificity were found to distinguish wild-type YF viruses from YF 17D-204 vaccine virus, but not from the closely related 17DD vaccine virus, nor from the French neurotropic vaccine virus. This pattern of reactivity was seen only with viruses grown in Aedes albopictus C6/36 cells and not with viruses grown in vertebrate cells (SW13 and Vero cells), where all five MAbs recognized epitopes on both wild-type and 17D-204 viruses. Examination of adult A. aegypti mosquitoes infected with the same YF viruses as above gave a different pattern of results to those in C6/36 cells. Thus, epitope expression differs between mammalian and arthropod cells and between arthropod cells in vitro and in vivo. Neutralization tests showed that all five MAbs would neutralize wild-type Asibi virus grown in SW13 cells, but not Asibi virus grown in C6/36 cells, nor 17D-204 vaccine virus grown in either cell type. Therefore, it is concluded that when YF virus is grown in mosquito cells, wild-type virus is antigenically and biologically distinct from the 17D-204 vaccine virus.

Examination of the envelope glycoprotein of yellow fever vaccine viruses with monoclonal antibodies.

Two panels of envelope glycoprotein reactive monoclonal antibodies (mAbs) were prepared against yellow fever (YF) 17D vaccine viruses. Five mAbs were prepared against the World Health Organization 17D-204 avian leukosis virus-free secondary seed virus and eight mAbs against 17DD vaccine manufactured in Brazil. The majority of these mAbs were type-specific and displayed differing reactions in neutralization tests. One, B14, would only neutralize YF vaccine virus grown in invertebrate cells. Others would differentiate 17D-204 and 17DD vaccines, from different manufacturers, in neutralization tests when the viruses were grown in vertebrate cells. The data indicate that heterogeneity exists between the epitopes that elicit neutralizing antibody on YF vaccine from different manufacturers.

Human monoclonal antibodies to thyroid antigens derived by hybridization of lymphocytes from a diabetic patient.

Human monoclonal antibodies to human endocrine cells have been obtained following the generation of immunoglobulin-secreting interspecies lymphocyte hybridomas. Peripheral blood lymphocytes from an adult patient presenting with acute onset, Type I, diabetes mellitus were fused in vitro with mouse myeloma cells of the NS1 cell line. Initial selection of resulting hybridomas was made by their ability to proliferate in HAT medium. Those hybridomas secreting human immunoglobulins were identified by radioimmunoassay and, thereafter, cloned at frequent intervals to ensure continued antibody production. Human monoclonal antibodies selected in this manner are being employed to identify those epitopes which are common antigenic targets during initial stages of autoimmune-mediated diabetes mellitus and associated multiple endocrinopathies. Of these antibodies, one (HML 3.22) recognizes an epitope present on the human TSH receptor and a second (HML 3.21) identifies a component of thyroglobulin. The potential value of human monoclonal antibodies as probes for analyzing autoimmune-mediated endocrine diseases is discussed.

The uptake and retrograde transport of horseradish peroxidase--polylysine conjugate by ligated postganglionic sympathetic nerves in vitro.

The uptake and retrograde transport of horseradish peroxidase (HRP) and horseradish peroxidase-poly-L-lysine conjugate (HRP-PL) were compared using a system comprising the guinea-pig inferior mesenteric ganglion (IMG) and ligated hypogastric hypogastric nerves maintained in vitro in a twin chamber apparatus. 0.5 mg of HRP-PL applied to the ligated nerves produced stronger retrograde labelling of neurons within the IMG than did 10 mg of HRP. This may have been due to the greater uptake of HRP-PL in a vesicular form by the axons immediately proximal to the ligation. The possible roles of large rounded vesicles and elongated cisternae in retrograde axoplasmic transport are discussed.

IGE in human urine and milk.

IgE was found in urine from healthy adult volunteers at very low levels (approximately 0.003-0.010 IU/ml), corresponding to a 24-h excrection rate of 3-16 IU. IgE was not detected in 36 out of 47 samples of milk and colostrum. In samples from six women the protein was present at low concentration 1-2 days postpartum but was not detected in later samples (usually day 5 or 6). Mammary secretions from four allergic donors were studied, and IgE was detected at low concentrations in samples from the two most severely affected individuals. The levels of IgE observed in both urine and milk suggest that there is no significant synthesis of the protein in either the urinary tract or in mammary tissue.