Clonal analysis of a human antibody response. Quantitation of precursors of antibody-producing cells and generation and characterization of monoclonal IgM, IgG, and IgA to rabies virus.

We quantitated and characterized the changes in the human B cell repertoire, at the clonal level, before and after immunization with rabies virus. Moreover, we generated 10 monoclonal cell lines producing IgM, IgG, and IgA antibodies to the virus. We found that in healthy subjects, not previously exposed to the virus, nearly 2% of the circulating B lymphocytes were committed to the production of antibodies that bound the virus. These B cells expressed the surface CD5 molecule. The antibodies they produced were polyreactive IgM that displayed a relatively low affinity for the virus components (Kd, 1.0-2.4 x 10(-6) g/microliters). After immunization, different anti-virus (IgG and IgA) antibody-producing cells consistently appeared in the circulation and increased from less than 0.005% to greater than 10% of the total B cells committed to the production of IgG and IgA, respectively. Most of such B cells do not express CD5 and produce monoreactive antibodies of high affinity for rabies virus (Kd, 6.5 x 10(-9) to 1.2 x 10(-10) g/microliters). One of these IgG mAbs efficiently neutralized rabies virus in vitro and in vivo, as detailed elsewhere (Dietzschold, B., P. Casali, Y. Ueki, M. Gore, C. E. Rupprecht, A. L. Notkins, and H. Koprowski, manuscript submitted for publication). Hybridization experiments using probes specific for the different human V gene segment families revealed that cell precursors producing low affinity IgM binding to rabies virus utilized a restricted number of VH gene segments (i.e., only members of the VHIIIb subfamily), whereas cell precursors producing high affinity IgG and IgA to rabies virus utilized an assortment of different VH gene segments (i.e., members of the VHI, VHIII, VHIV, and VHVI families and VHIIIb subfamily). In conclusion, our studies show that EBV transformation in conjunction with limiting dilution technology and somatic cell hybridization techniques are useful methods for quantitating, at the B cell clonal level, the human antibody response to foreign Ags and for generating human mAbs of predetermined specificity and high affinity

Juice constituents and skin pigments in Vitis coignetiae Pulliat grapevines

Research Note

Systematic Study of Short Range Antiferromagnetic Order and The Spin-Glass State in Lightly Doped La2-xSrxCuO4

Systematic measurements of the magnetic susceptibility were performed on single crystals of lightly doped La2-xSrxCuO4 (x=0.03, 0.04 and 0.05). For all samples the temperature dependence of the in-plane magnetic susceptibility shows typical spin-glass features with spin-glass transition temperatures Tg of 6.3K, 5.5K and 5.0K for x=0.03, 0.04 and 0.05, respectively. The canonical spin-glass order parameter extracted from the in-plane susceptibility of all the samples follows a universal scaling curve. On the other hand, the out-of-plane magnetic susceptibility deviates from Curie law below a temperature Tdv, higher than Tg. Comparing with previous neutron scattering results with an instrumental energy resolution of 0.25 meV from Wakimoto et al., the x-dependence of Tdv is qualitatively the same as that of Tel, the temperature below which the elastic magnetic scattering develops around (pi, pi). Thus, a revised magnetic phase diagram in the lightly doped region of La2-xSrxCuO4 is proposed. The Curie constants calculated from the in-plane susceptibility are independent of the Sr concentration. On the basis of the cluster spin-glass model, this fact might reflect an inhomogeneous distribution of doped holes in the CuO2 plane, such as in a stripe structure.Comment: 7 pages, 6 figure