Decrease of T-cells exhaustion markers programmed cell death-1 and T-cell immunoglobulin and mucin domain-containing protein 3 and plasma IL-10 levels after successful treatment of chronic hepatitis C

During chronic hepatitis C virus (HCV) infection, both CD4$^{+}$ and CD8$^{+}$ T-cells become functionally exhausted, which is reflected by increased expression of programmed cell death-1 (PD-1) and T-cell immunoglobulin and mucin domain-containing protein 3 (Tim-3), and elevated anti-inflammatory interleukin 10 (IL-10) plasma levels. We studied 76 DAA-treated HCV-positive patients and 18 non-infected controls. Flow cytometry measured pretreatment frequencies of CD4$^{+}$PD-1$^{+}$, CD4$^{+}$PD-1$^{+}$Tim-3$^{+}$ and CD8$^{+}$PD-1$^{+}$Tim-3$^{+}$ T-cells and IL-10 levels measured by ELISA were significantly higher and CD4$^{+}$PD-1$^{-}$Tim-3$^{-}$ and CD8$^{+}$PD-1$^{-}$Tim-3$^{-}$ T-cells were significantly lower in patients than in controls. Treatment resulted in significant decrease of CD4$^{+}$Tim-3$^{+}$, CD8$^{+}$Tim-3$^{+}$, CD4$^{+}$PD-1$^{+}$Tim-3$^{+}$ and CD8$^{+}$PD-1$^{+}$Tim-3$^{+}$ T-cell frequencies as well as IL-10 levels and increase in CD4$^{+}$PD-1$^{-}$Tim-3$^{-}$ and CD8$^{+}$PD-1$^{-}$Tim-3$^{-}$ T-cells. There were no significant changes in the frequencies of CD4$^{+}$PD-1$^{+}$ T-cells, while CD8$^{+}$PD-1$^{+}$ T-cells increased. Patients with advanced liver fibrosis had higher PD-1 and lower Tim-3 expression on CD4$^{+}$T-cells and treatment had little or no effect on the exhaustion markers. HCV-specific CD8$^{+}$T-cells frequency has declined significantly after treatment, but their PD-1 and Tim-3 expression did not change. Successful treatment of chronic hepatitis C with DAA is associated with reversal of immune exhaustion phenotype, but this effect is absent in patients with advanced liver fibrosis

T-Cell Exhaustion in HIV-1/Hepatitis C Virus Coinfection Is Reduced After Successful Treatment of Chronic Hepatitis C.

BACKGROUND T-cell responses during chronic viral infections become exhausted, which is reflected by upregulation of inhibitory receptors (iRs) and increased interleukin 10 (IL-10). We assessed 2 iRs-PD-1 (programmed cell death protein 1) and Tim-3 (T-cell immunoglobulin and mucin domain-containing protein 3)-and IL-10 mRNAs in peripheral blood mononuclear cells (PBMCs) and their soluble analogs (sPD-1, sTim-3, and IL-10) in plasma in chronic HIV-1/hepatitis C virus (HCV) coinfection and explored the effect of HCV treatment on these markers. We also aimed to establish whether iR expression may be determined by the HCV CD8+ T-cell immunodominant epitope sequence. METHODS Plasma and PBMCs from 31 persons with chronic HIV-1/HCV coinfection from the Swiss HIV Cohort Study were collected before and after HCV treatment. As controls, 45 persons who were HIV-1 negative with chronic HCV infection were recruited. Exhaustion markers were assessed by enzyme-linked immunosorbent assay in plasma and by quantitative reverse transcription polymerase chain reaction in PBMCs. Analysis of an HCV epitope sequence was conducted by next-generation sequencing: HLA-A*02-restricted NS31073-1081 and NS31406-1415 and HLA-A*01-restricted NS31436-1444. RESULTS The study revealed higher plasma sPD-1 (P = .0235) and IL-10 (P = .002) levels and higher IL-10 mRNA in PBMCs (P = .0149) in HIV-1/HCV coinfection. A decrease in plasma sPD-1 (P = .0006), sTim-3 (P = .0136), and IL-10 (P = .0003) and Tim-3 mRNA in PBMCs (P = .0210) was observed following successful HCV treatment. Infection with the HLA-A*01-restricted NS31436-1444 ATDALMTGY prototype variant was related to higher sTim-3 levels than infection with the ATDALMTGF escape variant (P = .0326). CONCLUSIONS The results underscore the synergistic effect of coinfection on expression of exhaustion markers, their reduction following successful HCV treatment and imply that iR levels may operate on an epitope-specific manner

T-Cell Exhaustion in HIV-1/Hepatitis C Virus Coinfection Is Reduced After Successful Treatment of Chronic Hepatitis C

BACKGROUND T-cell responses during chronic viral infections become exhausted, which is reflected by upregulation of inhibitory receptors (iRs) and increased interleukin 10 (IL-10). We assessed 2 iRs-PD-1 (programmed cell death protein 1) and Tim-3 (T-cell immunoglobulin and mucin domain-containing protein 3)-and IL-10 mRNAs in peripheral blood mononuclear cells (PBMCs) and their soluble analogs (sPD-1, sTim-3, and IL-10) in plasma in chronic HIV-1/hepatitis C virus (HCV) coinfection and explored the effect of HCV treatment on these markers. We also aimed to establish whether iR expression may be determined by the HCV CD8$^{+}$ T-cell immunodominant epitope sequence. METHODS Plasma and PBMCs from 31 persons with chronic HIV-1/HCV coinfection from the Swiss HIV Cohort Study were collected before and after HCV treatment. As controls, 45 persons who were HIV-1 negative with chronic HCV infection were recruited. Exhaustion markers were assessed by enzyme-linked immunosorbent assay in plasma and by quantitative reverse transcription polymerase chain reaction in PBMCs. Analysis of an HCV epitope sequence was conducted by next-generation sequencing: HLA-A*02-restricted NS3$_{1073-1081}$ and NS3$_{1406-1415}$ and HLA-A*01-restricted NS3$_{1436-1444}$. RESULTS The study revealed higher plasma sPD-1 (P = .0235) and IL-10 (P = .002) levels and higher IL-10 mRNA in PBMCs (P = .0149) in HIV-1/HCV coinfection. A decrease in plasma sPD-1 (P = .0006), sTim-3 (P = .0136), and IL-10 (P = .0003) and Tim-3 mRNA in PBMCs (P = .0210) was observed following successful HCV treatment. Infection with the HLA-A*01-restricted NS3$_{1436-1444}$ ATDALMTGY prototype variant was related to higher sTim-3 levels than infection with the ATDALMTGF escape variant (P = .0326). CONCLUSIONS The results underscore the synergistic effect of coinfection on expression of exhaustion markers, their reduction following successful HCV treatment and imply that iR levels may operate on an epitope-specific manner

Reversal of T cell exhaustion in chronic HCV infection

The long-term consequences of T cell responses' impairment in chronic HCV infection are not entirely characterized, although they may be essential in the context of the clinical course of infection, re-infection, treatment-mediated viral clearance and vaccine design. Furthermore, it is unclear whether a complete reinvigoration of HCV-specific T cell response may be feasible. In most studies, attempting to reverse the effects of compromised immune response quality by specific blockades of negative immune regulators, a restoration of functional competence of HCV-specific T cells was shown. This implies that HCV-induced immune dysfunction may be reversible. The advent of highly successful, direct-acting antiviral treatment (DAA) for chronic HCV infection instigated investigation whether the treatment-driven elimination of viral antigens restores T cell function. Most of studies demonstrated that DAA treatment may result in at least partial restoration of T cell immune function. They also suggest that a complete restoration comparable to that seen after spontaneous viral clearance may not be attained, pointing out that long-term antigenic stimulation imprints an irreversible change on the T cell compartment. Understanding the mechanisms of HCV-induced immune dysfunction and barriers to immune restoration following viral clearance is of utmost importance to diminish the possible long-term consequences of chronic HCV infection

Hepatitis C virus (HCV) genotype 1b displays higher genetic variability of hypervariable region 1 (HVR1) than genotype 3

Hepatitis C virus (HCV) is characterized by high genetic variability, which is manifested both at the inter-host and intra-host levels. However, its role in the clinical course of infection is less obvious. The aim of the present study was to determine the genetic variability of HCV HVR1 (hypervariable region 1) of genotype 1b and 3 in plasma of blood donors in the early seronegative stage of infection (HCV-RNA+, anti-HCV-) and in samples from chronically infected patients using next-generation sequencing. Sequencing errors were corrected, and haplotypes inferred using the ShoRAH software. Genetic diversity parameters (intra-host number of variants, number of nucleotide substitutions and diversity per site) were assessed by DNA SP and MEGA. During the early infection, the number of variants were significantly lower in subjects infected with genotype 3 than with genotype 1b (p < 0.02). Similarly, intra-host number of variants, number of nucleotide substitutions and diversity per site were lower in genotype 3 chronic infection (p < 0.0005). In addition, early infection was characterized by significantly lower HVR1 variability values (p < 0.04) when compared to chronic infection for both genotypes. It seems that the observed differences in HVR1 variability represent an inherent property of particular viral genotypes

The Beneficial Effect of the COVID-19 Vaccine Booster Dose among Healthcare Workers in an Infectious Diseases Center

Introduction: Healthcare workers in Poland received a booster dose of the BNT162b2 mRNA vaccine (Pfizer-BioNTech, Manufacturer: Pfizer, Inc., and BioNTech; Moguncja, Germany) at the beginning of October 2021. Here, we report on the preliminary results of an ongoing clinical study into the antibody response to SARS-CoV-2 of healthcare workers previously exposed to the virus, with or without evidence of past infection, in the Hospital for Infectious Diseases in Warsaw before and after the vaccine booster dose. Methods: Blood samples were collected on the day the vaccine booster dose was administered and again 14 days later. The levels of SARS-CoV-2 IgG antibodies (against the n-protein, indicative of disease) and S-RBD (indicative of a response to vaccination) were measured. Results: One hundred and ten health care workers from the Hospital for Infectious Diseases were included in the study. The percentage of subjects with a positive test for anti-n-protein IgG antibodies at both time points remained unchanged (16, 14%), while a statistically significant increase in the percentage of subjects producing high levels of S-RBD antibodies (i.e., >433 BAU/mL) was observed (from 23, 21% to 109, 99%; p = 0.00001). Conclusions: The results of the study indicate that the booster dose of the vaccine significantly increases the percentage of people with high levels of S-RBD antibodies, regardless of previous contact with the virus, which may indicate greater protection against both the disease and a severe course of COVID-19

The Beneficial Effect of the COVID-19 Vaccine Booster Dose among Healthcare Workers in an Infectious Diseases Center

Introduction: Healthcare workers in Poland received a booster dose of the BNT162b2 mRNA vaccine (Pfizer-BioNTech, Manufacturer: Pfizer, Inc., and BioNTech; Moguncja, Germany) at the beginning of October 2021. Here, we report on the preliminary results of an ongoing clinical study into the antibody response to SARS-CoV-2 of healthcare workers previously exposed to the virus, with or without evidence of past infection, in the Hospital for Infectious Diseases in Warsaw before and after the vaccine booster dose. Methods: Blood samples were collected on the day the vaccine booster dose was administered and again 14 days later. The levels of SARS-CoV-2 IgG antibodies (against the n-protein, indicative of disease) and S-RBD (indicative of a response to vaccination) were measured. Results: One hundred and ten health care workers from the Hospital for Infectious Diseases were included in the study. The percentage of subjects with a positive test for anti-n-protein IgG antibodies at both time points remained unchanged (16, 14%), while a statistically significant increase in the percentage of subjects producing high levels of S-RBD antibodies (i.e., &gt;433 BAU/mL) was observed (from 23, 21% to 109, 99%; p = 0.00001). Conclusions: The results of the study indicate that the booster dose of the vaccine significantly increases the percentage of people with high levels of S-RBD antibodies, regardless of previous contact with the virus, which may indicate greater protection against both the disease and a severe course of COVID-19

Wyniki wieloośrodkowych badań dotyczących diagnostyki i leczenia raka krtani w Polsce w latach 2001–2010

Wstęp: Rak krtani jest w Polsce najczęściej występującym nowotworem złośliwym regionu głowy i szyi. Rozpoznaje się go przede wszystkim u mężczyzn pomiędzy 50.–70. rokiem życia. Badania wieloośrodkowe, których celem było przedstawienie obrazu epidemiologicznego i klinicznego raka krtani, prowadzone były od 1980 roku. Obecne, koordynowane przez Klinikę Otolaryngologii WUM, mają za główny cel przedstawienie ewolucji procesu diagnostycznego i terapeutycznego. Materiał i metody: Analizie retrospektywnej poddano dane dotyczące chorych diagnozowanych i leczonych z powodu raka krtani w latach 2001–2010 w 12 ośrodkach. Na platformie bazy Microsoft Access 2003 (SP 2) stworzono program do gromadzenia danych, które następnie poddano analizie statystycznej. Wyniki: Zgromadzono dane dotyczące 4124 chorych, w tym 3682 mężczyzn (89,3%) i 442 kobiet (10,7%). Najliczniejszą grupę stanowili chorzy pomiędzy 50. a 60. rokiem życia – 41,5% oraz między 60. a 70. rokiem życia – 29,6%. Aż 81,3% pacjentów było nałogowymi palaczami papierosów. U 1634 chorych stwierdzono I i II stopień zaawansowania nowotworu, przy czym u 5 chorych był to rak in situ. U 2490 chorych zdiagnozowano III i IV stopień zaawansowania nowotworu. Najczęstszym umiejscowieniem raka były: głośnia, trzy piętra krtani oraz nadgłośnia i głośnia. Największą grupę stanowili pacjenci z rakiem krtani T3 – 1367 (33%). Węzły chłonne (cecha N) obecne były u 1216 chorych (29,5%). Największą grupę stanowiły węzły określane jako N2b i N2c, czyli powyżej 6 cm, oraz mnogie. Zdiagnozowano je u 533 chorych (13%). Węzły te występowały z rakami krtani określanymi jako T3 i T4a, a więc były to nowotwory zaawansowane miejscowo i regionalnie. Rodzaj zastosowanego leczenia bardzo różnił się w zależności od stopnia zaawansowania nowotworu. Sama chirurgia dotyczyła 73,7% pacjentów z nowotworem w stopniu I lub II, oraz jedynie 28,6% pacjentów z nowotworem w stopniu III lub IV. Nowotwory w zaawansowanych stadiach znacznie częściej były leczone terapią łączoną: chirurgią i radioterapią. Wyniki leczenia – mierzone przeżyciem całkowitym – wynosiły odpowiednio: w grupie chorych z zawansowaniem raka w I i II stopniu – 64%, w grupie chorych z zaawansowaniem raka w stopniu III i IV – 61%. Wnioski: Skuteczność rozpoznawania raka krtani w Polsce jest duża, jednak skróceniu powinien ulec czas upływający od wystąpienia u pacjenta objawów do uzyskania diagnozy. Natomiast skuteczność leczenia chirurgicznego jest zdecydowanie niezadowalająca. Stworzenie stałej platformy do prowadzenia (i monitorowania) przebiegu diagnostyki i leczenia raka krtani, pozwoli na zweryfikowanie postępowania i osiągnięcie lepszych wyników. Jest to zadanie dla środowiska otorynolaryngologów

The multicenter study result of diagnosis and treatment laryngeal carcinoma in Poland from 2001 to 2010

Introduction: The laryngeal cancer is the most frequently diagnose malignancy in head and neck region. The highest morbidity is within male patients in the age range between 50 and 70 years. The multicenter study, coordinated by Oto-laryngology Department of Medical University of Warsaw, was designed to investigate the epidemiology of laryngeal cancer in Poland from 1980 and to analyze the evolution of diagnostic and therapeutical procedures over the years.Material and methods: There was performed retrospective analysis of the medical records of patients with laryn-geal cancer hospitalized and treated in 12 otolaryngology centers in Poland from 2001 to 2010. The Microsoft Access 2003 (SP 2) platform was used to collect the data and subsequent statistical analysis.Results: There were collected data from 4124 patients, 3682 men (89,3%) and 442 women (10,7%). The largest group consisted of patients in the age range between 50 and 60 years (41,5%) and the second large group was of those be-tween 60 and 70 years (29,6%). The history of heavy cigarettes smoking was obtained from 81,3% of patients. Re-garding the staging of laryngeal cancer, there were 1634 patients with cancer stage of I or II, including 5 patients with carcinoma in situ and 2490 patients with III or IV stage. The most frequent localization of the cancer was the glottis, followed with invasion of all three laryngeal levels and tumors occupying both the epiglottis and glottis. The major-ity of patients – 1367 (33%) – has the T3 tumor advancement. The lymph nodes metastases (N) were present in 1216 (29,5%) patients and among them the N2b and N2c advancement ( lymph node larger than 6 cm, multiple) was de-tected in 533 of patients (13%). The lymph nodes involvement occurred in majority within advanced tumors of T3 or T4a. Considering the treatment options they varied depending on the staging of laryngeal cancer. The sole surgical procedures were performed in 73,7% of patients with cancer stage of I or II and in only 28.6% of patients with the stage of III or IV. The advanced tumors were in majority treated with the combined therapy: surgery and radiotherapy. The overall survival in patients with cancer stage of I and II was 64% and 61% for those with stage III and IV.Conclusions: The rate of laryngeal cancer detection is quite high in Poland, however the period from the onset of symptoms until diagnosis should be reduced. The effectiveness of surgical treatment is definitely unsatisfactory. The project to create a multicenter permanent base for monitoring the course of diagnosis and treatment in patients with laryngeal cancer surely will verify the procedures and enable to achieve better results. Continuation of this project is a task for all otorhinolaryngologists.KEYWORDS:laryngeal carcinoma, epidemiology, clinical characteristics, multicenter stud

Highlighter plot showing differences in amino acid composition of HVR1 HCV sequence variants in hospitalized patients from the infection cluster.

<p>Variants are compared to consensus sequence built from all sequences present in all patients. Color coding of amino acid is shown in the legend. Potential N-glycosylation site in the consensus sequence is marked by a purple dot. Amino acid numbering follows the reference strain H77 (GenBank accession no. AF009606). HVR1 spans codons 384–410.</p