7 P De Togn e t a / , i b~d , p 703, T. A Banks et a1 , J /n?n?uno/ 155, 1685 (1 995) 8 Mce were ~mmun~zed intrapertoneally w~th 100 &I of phosphate-buffered sane contanng 10% SRBCs (22) Ten days later, spleens were harvested, embedded In 0 C.T compound (M~les), and frozen In q u d ntrogen Frozen t~ssue sections (6 to 10 &m th~ck) were f~xed n cold acetone Endogenous per-ox~dase was q~~enched w~th 0 2% H, O, n methanol Sectons were sta~ned by f~rst ncubatng w~t h PNA-b~o t~n (Vector) and w~th rat antserum produced to IgD (Southern B~otechnology) After wash~ng, the sections were further incubated w~th strepta\/id~n conj~lgated w~th alkalne phosphatase (AP) (Zymed) and rabb~t ant~body to rat IgG conjugated w~th horse-rad~sh perox~dase (HRP) (Southern B~otechnology) Color development for bound AP and HRP was w~th an AP reacton k~t (Vector) and w~t h d~arn~nobenz-d~ne Sect~ons were then countersta~ned w~th 1 % methyl green 9 M. L Rose, M S C B~rbeck, V J Wall~s, J A Forrester. A J Daves. Nature 284, 364 (1980) 10 T

FPRL1 and GPR1 have the potential to work as significant HIV-1 coreceptors in vivo next to CCR5 and CXCR4. HIV-1 populations that can use various GPCRs as coreceptors are already circulating in vivo, even in the early stage of HIV-1 infection.

In this study, the first complete genome sequences for hepatitis C virus (HCV) subtypes 6f, 6i, 6j and 6m, obtained from infected blood donors in Chiang Mai, Thailand, are reported. Pairwise genome-wide nucleotide similarities between some of these isolates were higher than the 75-80 % value used previously to define different HCV subtypes. To investigate further, the entire genomes of four prototype isolates, Th602 (6i), Th553 (6j), B4/92 (6m) and D86/93 (6n), were sequenced. Pairwise comparison of these sequences gave a similar range of nucleotide similarities, thereby providing new information for HCV subtype classification. In order to study the hypothesis of interspousal HCV transmission, four additional complete HCV genome sequences were obtained from two infected Thai blood donors and their spouses, C-0044 and C-0046 (6f), and C-0192 and C-0185 (6m). Pairwise comparison of the sequences revealed that C-0044 and C-0046 share a nucleotide similarity of 98.1 %, whilst C-0185 and C-0192 have a similarity of 97.8 %. Several other studies of partial HCV sequences of different genomic regions from HCV-infected couples have shown nucleotide similarities ranging from 96.3 to 100 %. The similarities of the complete genome sequences from the two couples in the current study are consistent with HCV transmission between spouses.

Subtype analysis of hepatitis C viruses (HCVs) obtained from patients with chronic liver disease in ChiangMai, Thailand, was performed. Of 46 HCV isolates, 13 (28%) were shown to belong to HCV subtype 3a (HCV-3a), 10 (22%) to belong to HCV-1a, 7 (15%) to belong to HCV-1b, 1 (2%) to belong to HCV-3b, and 1 (2%) to belong to a variant group, as determined from partial nucleotide sequences of the NS5B region of the viral genome. Analysis of 5 untranslated region sequences identified five other isolates (11%) of HCV type 1 and two other isolates (4%) of type 3. Detailed phylogenetic positions for the variant described above and those previously obtained from blood donors and drug addicts in Chiang Mai were determined by a six-parameter neighbor-joining method on the basis of core, E1, and NS5B region sequences. The results revealed that those sequence variants represent novel subtypes of HCV type 6. The HCV type 6 isolates appear to be antigenically different from isolates of HCV types 1 and 2, as determined by a serotyping method that utilizes recombinant peptides corresponding to a portion of the NS4 protein. The significance of subtype analysis around this area is discussed.Hepatitis C virus (HCV) has been known to be a major etiologic agent of posttransfusion as well as sporadic non-A, non-B hepatitis worldwide (14). The genome of HCV is a single-stranded RNA with positive polarity of about 9,400 bases, which has a long open reading frame encoding four structural proteins (core, E1, E2 type A, and E2 type B) and at least six nonstructural proteins (NS2, NS3, NS4A, NS4B, NS5A, and NS5B) with untranslated regions at both the 5Ј and 3Ј ends (5ЈUTR and 3ЈUTR, respectively) (8, 10, 18). The viral genome exhibits considerable degrees of sequence diversity among different isolates. On the basis of this diversity, HCV has been classified into a number of distinct types or groups, each of which can be further divided into several subtypes (4, 24-26). Viral pathogenicity and susceptibility to interferon treatment appear to vary among different subtypes (15,21,22). Also, the prevalence of each subtype among HCV isolates varies among geographical areas (5,9,25). Moreover, the identification of sequence variants that might represent novel types or subtypes has been increasing as surveillance is extended to previously overlooked areas (1, 4, 11-13, 20, 28, 29).We previously determined the prevalence of each subtype among HCV isolates from anti-HCV-positive blood donors and drug addicts in Chiang Mai, Thailand, and identified a number of unique sequence variants (1). However, the exact phylogenetic positions of the variants could not be determined at that time. In the present study we have determined the HCV subtype prevalence among anti-HCV-positive patients with chronic liver disease. We also performed a detailed phylogenetic analysis that shows that the sequence variants obtained in this area represent novel subtypes of HCV type 6. We discuss how the variants are likely to possess an antigenic epitope(s) that is distinct fro...

Available data suggest that of the currently available options, treatment of TE with pyrimethamine at 50 mg/day plus sulfadiazidine at 4 g/day provides the best primary outcome for AIDS patients with TE; however, because this study was terminated prematurely, we suggest that treatment with intravenous TMP-SMX be further evaluated to determine its efficacy.