109 research outputs found
Clonal analysis of intrahepatic T lymphocytes in chronic active hepatitis. Isolation of a T-cell line specific for hepatitis B core antigen from a patient with serological evidence of exposure to HBV
No full textWe have evaluated whether peripheral blood and hepatic lymphocytes from a patient with chronic active hepatitis (CAH) and antibodies to HBV in serum were specifically sensitized to HBV envelope antigens (HBsAg and pre-S Ag) or to HBcAg. No proliferation to HBV antigens was demonstrated upon stimulation of peripheral blood mononuclear cells either unfractionated or enriched in CD4+ (helper/inducer) T cells. Of 15 T-cell cloned lines (7 CD8+ and 8 CD4+) obtained by limiting dilution in the presence of PHA and recombinant IL2 from liver-infiltrating lymphocytes, one, designated H2, showed specific sensitization to HBcAg, whereas none demonstrated sensitization to viral envelope antigens. The H2 line displayed the CD8+ phenotype, suppressor activity on polyclonal immunoglobulin production and IL2-dependent, HBcAg-specific proliferation. These results suggest that in patients with CAH and serological evidence of previous exposure to HBV, it is possible to obtain lymphocytes specifically sensitized to HBcAg from liver biopsy
Intrahepatic, nucleocapsid antigen-specific T cells in chronic active hepatitis B
No full textHepatitis B core antigen (HBcAg)-specific T cell lines were established from hepatic lymphomononuclear cells derived from five patients with chronic active hepatitis B. No hepatitis B virus envelope antigen-specific cell lines were established. Proliferation in response to recombinant and native HBcAg, but not to native hepatitis B surface antigen containing the pre-S(2) region, confirmed the specificity of the five T cell lines. All cell lines represented mixed populations of CD4+ and CD8+ T cells. The CD4+ subset provided antigen-specific help to autologous B cells with respect to anti-HBc production and to CD8+ cells with regard to HBcAg-induced proliferation and suppressor activity. The CD8+ subset contained suppressor cells that selectively inhibited the proliferative response of autologous HBcAg-specific CD4+ cells without inhibiting CD4+ cells of unrelated specificity (tetanus toxoid). Moreover, the CD8+ cells were also capable of suppressing HBcAg-stimulated antibody to HBcAg production without showing inhibition of total immunoglobulin production stimulated by pokeweed mitogen. The cytotoxic potential of the T cell lines was established in a lectin-dependent cytotoxicity system; natural killer cytotoxicity was completely absent. Our data suggest that the lesional T cells present at the site of hepatocellular injury in chronic active hepatitis B are primarily HBcAg-specific lymphocytes of the helper and suppressor/cytotoxic phenotypes and that both are functionally competent
Selective sensitization of peripheral blood T lymphocytes to hepatitis B core antigen in patients with chronic active hepatitis type B.
No full textThe proliferative response of peripheral blood T cells to hepatitis B surface antigen (HBsAg), pre-S antigen and hepatitis B core antigen (HBcAg) has been studied in 20 patients with hepatitis B virus induced chronic active hepatitis (CAH) and in 12 control subjects. Eleven of the 20 CAH patients showed a significant T lymphocyte proliferative response to HBcAg, whereas no proliferation was detectable in response to envelope antigens (HBs and pre-S) in any patient. T cell subset fractionation revealed that HBcAg specific proliferation was limited to the CD4+ (helper/inducer) population. CD8+ (suppressor/cytotoxic) T cells were unresponsive to HBcAg and did not suppress the proliferative response of autologous CD4+ cells. The HBcAg specific T cell proliferative response did not correlate with serum anti-core antibody titres or with biochemical evidence of liver disease. The present results show the selective presence of HBcAg-sensitized T cells in the peripheral blood of patients with HBsAg positive CAH and suggest that the peripheral lymphoid compartment may not reflect immunopathogenetically important cellular events operative at the site of tissue injur
Clonal analysis of intrahepatic T lymphocytes in chronic active hepatitis. Isolation of a T-cell line specific for hepatitis B core antigen from a patient with serological evidence of exposure to HBV. J. Hepatol.
No full textRestored Function of HBV-Specific T Cells After Long-term Effective Therapy With Nucleos(t)ide Analogues.
No full textBACKGROUND & AIMS:
In patients with chronic hepatitis B virus (HBV) infection, persistent exposure to high concentrations of antigen can disrupt T-cell functions. It is not clear to what extent long-term suppression of HBV by nucleos(t)ide analogues can restore antiviral T-cell functions. We compared HBV-specific T-cell responses of patients treated with nucleos(t)ide analogues with those detected in other conditions of HBV control.
METHODS:
We analyzed intracellular levels of interferon gamma, interleukin-2, and tumor necrosis factor α in HBV-specific T cells after 10 days of stimulation with peptides covering the overall HBV genotype D sequence and ex vivo with selected CD8 epitopes and the corresponding HLA-A2 dextramers. Findings from patients treated with nucleos(t)ide analogues who had complete (HBV DNA negative/antibody to hepatitis B surface antigen positive) or partial (HBV DNA negative/hepatitis B surface antigen positive) control of their infections were compared with those of patients with spontaneous or interferon alfa-induced resolution of acute or chronic infections, inactive HBV carriers, or untreated hepatitis B e antigen-negative patients with chronic infections.
RESULTS:
Although HBV-specific T cells from nucleos(t)ide analogue-treated patients with complete control of infection were dysfunctional ex vivo, they had efficient responses after in vitro expansion. These responses were comparable to those of patients who spontaneously resolved acute HBV infection. Nucleos(t)ide analogue-treated patients who were HBV DNA negative but hepatitis B surface antigen positive had lower levels of T-cell responses but responses greater than those of untreated patients with chronic infection.
CONCLUSIONS:
In vitro reactivity can be restored to T cells from patients with suppressed HBV infection following long-term treatment with nucleos(t)ide analogues, despite prolonged exposure to large loads of antigen. Immune therapies that increase the antiviral T-cell response might increase the likelihood of complete HBV control in patients undergoing long-term nucleos(t)ide analogue treatment.
Copyright © 2012 AGA Institute. Published by Elsevier Inc. All rights reserve
Technical note: Quantification of zeins from corn, high-moisture corn, and corn silage using a turbidimetric method: Comparative efficiencies of isopropyl and tert-butyl alcohols
No full textZeins are corn endosperm storage proteins that encapsulate
starch granules into a protein matrix, which
can act as a barrier to starch accessibility and digestion.
Laboratory methods to quantify zein are seldom
used because they are considered arduous and timeconsuming.
A recently published rapid turbidimetric
method (mTM) was reinvestigated by changing the
solution originally used for the zein solubilization step.
In particular, the aim was to explore whether, and to
what extent, the use of tert-butyl alcohol (t-BuOHmTM)
in lieu of isopropyl alcohol (i-PrOH-mTM) was
able to improve the quantification of zeins from dry
corn, high-moisture corn, and corn silage samples. The
nature of the alcohol influenced the zein extraction
values, and t-BuOH-mTM gave higher zein values in
corn (3.6 vs. 3.3 g/100 g of dry matter) and corn silage
samples (1.2 vs. 0.9 g/100 g of dry matter) compared
with i-PrOH-mTM. In contrast, similar zein extraction
values were obtained for high-moisture corn (2.1 vs.
1.9 g/100 g of dry matter, respectively). Sodium dodecyl
sulfate-PAGE analysis revealed no contamination
by nonzein proteins with the use of tert-butyl alcohol.
Overall, these findings indicated that tert-butyl alcohol
has a greater ability to solubilize zein compared with
isopropyl alcohol and thus the t-BuOH-mTM allowed
greater extraction of zeins. Considering the growing
interest of animal nutritionists in zein proteins, such
results should provide useful informatio
Cytotoxic T lymphocytes recognize an HLA-A2-restricted epitope within the hepatitis B virus nucleocapsid antigen.
No full textThe absence of readily manipulable experimental systems to study the cytotoxic T lymphocyte (CTL) response against hepatitis B virus (HBV) antigens has thus far precluded a definitive demonstration of the role played by this response in the pathogenesis of liver cell injury and viral clearance during HBV infection. To circumvent the problem that HBV infection of human cells in vitro for production of stimulator/target systems for CTL analysis is not feasible, a panel of 22 overlapping synthetic peptides covering the entire amino acid sequence of the HBV core (HBcAg) and e (HBeAg) antigens were used to induce and to analyze the HBV nucleocapsid-specific CTL response in nine patients with acute hepatitis B, six patients with chronic active hepatitis B, and eight normal controls. By using this approach, we have identified an HLA-A2-restricted CTL epitope, located within the NH2-terminal region of the HBV core molecule, which is shared with the e antigen and is readily recognized by peripheral blood mononuclear cells from patients with self-limited acute hepatitis B but less efficiently in chronic HBV infection. Our study provides the first direct evidence of HLA class I-restricted T cell cytotoxicity against HBV in humans. Furthermore, the different response in HBV-infected subjects who successfully clear the virus (acute patients) in comparison with patients who do not succeed (chronic patients) suggests a pathogenetic role for this CTL activity in the clearance of HBV infection
Fine specificity of the human T cell response to hepatitis B virus core antigen
No full textThe fine specificity of the human T cell response to the hepatitis B virus core antigen (HBcAg) was investigated in 23 patients with acute hepatitis B virus (HBV) infection using a panel of short synthetic peptides covering the entire core region. An immunodominant T cell epitope which was recognized by all except one patient, was identified within the core sequence 50-69. Two further important T cell recognition sites were represented by the amino acid sequences 1-20 and 117-131, which were stimulatory for the T cells of 69% and 73% of the patients, respectively. T cell recognition of the synthetic peptides was HLA class II restricted because the peptide-induced T cell proliferation was inhibited by anti-HLA class II but not by anti-HLA class I monoclonal antibodies. These findings may be relevant to the development of future preventive and therapeutic strategies against HBV infection
Selective sensitization of peripheral blood T lymphocytes to hepatitis B core antigen in patients with chronic active hepatitis type B.
No full textFine specificity of the human T-cell response to the hepatitis B virus preS1 antigen.
No full textThe T-cell response to hepatitis B virus envelope antigens was studied in 11 hepatitis B vaccine recipients; 7 were selected to analyze the fine specificity of the T-cell response to the preS1 antigen. Four distinct T-cell epitopes were identified by peripheral blood lymphomononuclear cell stimulation with a panel of short synthetic peptides covering the preS1 sequence. The immunodominance of the preS1 epitopes included within peptides 21-30 and 29-48 was shown by their capacity to restimulate an HLA class II restricted proliferative response of T cells primed with the whole preS1 antigen. Conversely, peptide-specific T cells selected by peripheral blood lymphomononuclear cell stimulation with peptides 21-30 and 29-48 were able to recognize the native preS1 molecule, confirming that these epitopes are actually generated by the intracellular processing of preS1. Finally, amino acid residues essential for T-cell activation by peptide 21-30 were identified using 10 analogues of the stimulatory peptide containing single alanine substitutions. These results may be relevant to the design of efficient synthetic vaccines against hepatitis B virus infection
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