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Background & Aims: The immune response of gastric T cells during acute Helicobacter pylori infection has not been previously characterized. The aim of this study was to delineate the phenotypic and functional responses of gastric T cells during acute H. pylori infection of rhesus macaques. Methods: Four monkeys were experimentally infected with H. pylori. Gastric biopsy specimens and peripheral blood samples were obtained 1 and 12 weeks after inoculation. Samples from 3 animals uninfected with H. pylori served as controls. The immunophenotypic changes and functional potential of CD4^+ and CD8^+ T cells in gastric mucosa and peripheral blood to produce cytokines (interleukin [IL]-2, IL-4, IL-13, interferon [IFN]-@c, MIP-1@b, and tumor necrosis factor [TNF]-@a) were determined at a single cell level using flow cytometry. Results: An increase in CD4^+ T cells occurred in the gastric mucosa during acute H. pylori infection as early as 1 week after infection. Acute infection was characterized by a predominantly T helper (Th)1 (IL-2 and IFN-@c) and proinflammatory (TNF-@a and MIP-1@b) type of cytokine response and the absence of a Th2 type of response. Conclusions: A predominant Th1 type response was induced early during acute H. pylori infection and may contribute to the development of gastric disease. GASTROENTEROLOGY 2000;118:307-315

Using the simian immunodeficiency virus (SIV)-infected rhesus macaque model, we performed a longitudinal study to determine the effect of antiretroviral therapy on the phenotype and functional potential of CD4(+) T cells repopulating intestinal mucosa in human immunodeficiency virus infection. Severe depletion of CD4(+) and CD4(+) CD8(+) T cells occurred in the intestinal mucosa during primary SIV infection. The majority of these cells were of activated memory phenotype. Phosphonate 9-[2-(phosphomethoxypropyl]adenine (PMPA) treatment led to a moderate suppression of intestinal viral loads and repopulation of intestinal mucosa by predominantly activated memory CD4(+) T-helper cells. This repopulation was independent of the level of viral suppression. Compared to preinfection values, the frequency of naive CD4(+) T cells increased following PMPA therapy, suggesting that new CD4(+) T cells were repopulating the intestinal mucosa. Repopulation by CD4(+) CD8(+) T cells was not observed in either jejunum or colon lamina propria. The majority of CD4(+) T cells repopulating the intestinal mucosa following PMPA therapy were CD29(hi) and CD11ahi. A subset of repopulating intestinal CD4(+) T cells expressed Ki-67 antigen, indicating that local proliferation may play a role in the repopulation process. Although the majority of repopulating CD4(+) T cells in the intestinal mucosa were functionally capable of providing B- and T-cell help, as evidenced by their expression of CD28, these CD4(+) T cells were found to have a reduced capacity to produce interleukin-2 (IL-2) compared to the potential of CD4(+) T cells prior to SIV infection. Persistent viral infection may play a role in suppressing the potential of repopulating CD4(+) T cells to produce IL-2. Hence, successful antiretroviral therapy should aim at complete suppression of viral loads in mucosal lymphoid tissues, such as intestinal mucosa.

Genomic DNAs from dimethylbenzanthracene-induced BALB/c mouse mammary tumors arising from the transplantable hyperplastic outgrowth (HPO) line designated DI/UCD transformed NIH 3T3 cells upon transfection. Transforming activity was attributed to the presence of activated Harvey ras-1 oncogenes containing an A----T transversion at the middle adenosine nucleotide in codon 61. DNAs from untreated DI/UCD HPO cells and radiation-induced and spontaneous mammary tumors from the DI/UCD HPO line failed to transform NIH 3T3 cells. The results indicated that the mutation activation of Harvey ras-1 oncogenes was specific to dimethylbenzanthracene treatment in the mouse mammary tumor system.

The biosynthesis of the protein precursor of [Met]enkephalin (Tyr-Gly-Gly-Phe-Met) was studied using cell-free translation systems to characterize the enkephalin-precursor gene product and mRNA. Proteins obtained by translation of bovine adrenal medullary mRNA in the presence of [35S]methionine were digested with trypsin and carboxypeptidase B. The resultant peptides were immunoprecipitated with anti[Met]enkephalin serum and subsequently analyzed by reverse phase HPLC. [35S][Met]Enkephalin (up to 0.2 fmol/micrograms mRNA, identified by antigenic specificity and chromatographic mobility, was isolated from peptides obtained from proteins whose synthesis was dependent upon adrenal medullary mRNA. Both trypsin and carboxy-peptidase B were required to generate [35S][Met]enkephalin from translation products. The largest protein containing the [35S]-[Met]enkephalin sequence synthesized in the wheat germ system has a Mr, of 31,000 +/- 1000, determined by NaDodSO4/polyacrylamide gel electrophoresis. Adrenal medullary mRNA coding for protein containing the [35S][Met]enkephalin sequence was resolved into a major fraction of 1450 +/- 150 nucleotides and a minor fraction of 47000 +/- 450 nucleotides, as determined by agarose gel electrophoresis in the presence of methylmercuric hydroxide. It is proposed that the major enkephalin-precursor gene product of adrenal medulla is a protein of Mr approximately 31,000.U

The biosynthesis and initial processing of the methionine-enkephalin precursor preproenkephalin A were examined by cell-free translation of mRNA from brain and adrenal medulla. A novel antiserum raised against Met-enkephalin-Arg6-Phe7 was shown to react with bovine adrenal medulla fractions (apparent Mr 34,000) containing proenkephalin A. Affinity-purified antibodies from this antiserum were used to immunoprecipitate cell-free translated [35S]Met-enkephalin-containing protein. A protein of apparent Mr 30,000 +/- 500 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was the only Met-enkephalin precursor consistently synthesized by translation of mRNA from bovine or guinea pig striatum, rat brain, or bovine adrenal medulla. The presence of [35S]Met-enkephalin sequences in this protein was confirmed by high pressure liquid chromatography of trypsin/carboxypeptidase B digests. Dog pancreas endoplasmic reticulum membranes converted the Mr 30,000 protein to an immunoreactive protein of apparent Mr 28,500 that lacked significant core glycosylation. These results suggest that 1) a protein similar or identical to bovine adrenal medullary preproenkephalin A is the major Met-enkephalin precursor synthesized in brain as well as adrenal medulla, and 2) preproenkephalin A is converted to a protein resembling proenkephalin A, presumably by removal of a signal peptide.

Abstract. The presence of an immunoreactive thyroglobulin-like material in saliva from normal subjects and from 35 patients with thyroid carcinoma was detected by radioimmunoassay. The levels of this material in saliva were markedly elevated in patients with extensive metastases.Concentrated saliva samples from normal subjects and from patients with thyroid carcinoma were fractioned on Sepharose-6B and each fraction was assayed for thyroglobulin content by RIA. Several protein peaks of varying molecular size with thyroglublin-like immunoreactivity were observed. The physiological significance of these molecules in saliva remains to be established.

Serum thyroglobulin (tg) levels were measured in 112 patients with proved differentiated thyroid carcinoma (6 pre-operative, 32 post-operative and 74 in whom thyroidectomy was followed by radioiodine treatment). The tg levels were within the normal range in patients who had a residual mass in the neck region after surgery and who were in remission. The patients with metastases showed a wide range (undetectable to microgram quantities) of tg in circulation. The analysis of serum tg with respect to histology showed that the follicular variety of carcinoma tends to show higher tg in serum than the papillary type.Metastatic involvement of bone was associated with much higher serum tg levels than other sites such as lung and lymph node.Thirteen patients with bone metastases of non-thyroidal primary origin had tg levels within the normal range. Serum tg estimation is useful in excluding thyroid as a primary site in patients with bone metastases of unknown primary origin.

A dominant restriction allele, Akvr-1r, from California wild mice (Mus musculus domesticus) confers resistance to exogenous ecotropic murine leukemia virus (MuLV) infection. The presence of an ecotropic MuLV envelope-related glycoprotein in uninfected virus-resistant cells suggests that viral interference is a possible mechanism for this resistance. We molecularly cloned the ecotropic MuLV envelope-related sequence from the genomic DNA of a wild mouse homozygous for the Akvr-1r locus. The cloned provirus was defective and contained a C-terminal end of the pol gene, a complete envelope gene, and a 3' long terminal repeat. The presence of this provirus was directly correlated with Akvr-1r-mediated virus resistance in cell cultures and hybrid mice. The Akvr-1r provirus restriction map and partial DNA sequence were identical to those of the Fv-4r allele, an ecotropic MuLV resistance locus from Japanese feral mice (M. musculus molossinus), which was previously shown to be allelic with the Akvr-1r gene. The 3' host flanking sequences of Fv-4r and Akvr-1r also had identical restriction maps. These findings indicate that Akvr-1r and Fv-4r are the same gene. It was probably acquired by interbreeding of these feral species in recent times. Conservation of this locus might be favored by the useful function that it performs in protection against ecotropic MuLV infection endemic in both populations of wild mice.