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Highlights • Lymphopenia and eosinopenia might be predictors of disease severity in COVID-19. • Lymphopenia and eosinopenia might be predictors of disease progression in COVID-19. • Clinical classification-severe type is the dependently risk factor for alteration of PBICs. • PBICs might be a sentinel, and it deserves attention during COVID-19 management., Background The underlying changes of peripheral blood inflammatory cells (PBICs) in COVID-19 patients are little known. Moreover, the risk factors for the underlying changes of PBICs and their predicting role in severe COVID-19 patients remain uncertain. Material and Methods This retrospective study including two cohorts: the main cohort enrolling 45 patients of severe type serving as study group, and the secondary cohort enrolling 12 patients of no-severe type serving as control group. The PBICs analysis was based on blood routine and lymphocyte subsets. The inflammatory cell levels were compared among patients according to clinical classifications, disease-associated phases, as well as one-month outcomes. Results Compared with patients of non-severe type, the patients of severe type suffered from significantly decreased counts of lymphocytes, eosinophils, basophils, but increased counts of neutrophils. These PBICs alterations got improved in recovery phase, but persisted or got worse in aggravated phase. Compared with patients in discharged group, the patients in un-discharged/died group suffered from decreased counts of total T lymphocytes, CD4+T lymphocytes, CD8+T lymphocytes, as well as NK cells at 2 weeks after treatment. Clinical classification-critically severe was the independently risk factor for lymphopenia (OR=7.701, 95%CI:1.265-46.893, P=0.027), eosinopenia (OR=5.595, 95%CI:1.008-31.054, P=0.049) and worse one-month outcome (OR=8.984; 95%CI:1.021-79.061, P=0.048). Conclusion Lymphopenia and eosinopenia may serve as predictors of disease severity and disease progression in COVID-19 patients, and enhancing the cellular immunity may contribute to COVID-19 treatment. Thus, PBICs might become a sentinel of COVID-19, and it deserves attention during COVID-19 treatment.

Highlights • We investigate the dynamic changes of peripheral blood lymphocytes subsets in adult patients with COVID-19. • The levels of peripheral blood lymphocyte subsets (CD3 +, CD4 +, and CD8 +) are significantly different between non-severe and severe patients with COVID-19. • The levels of peripheral blood lymphocyte subsets (CD3 +, CD4 +, and CD8 +) are significantly different between recovered and death patients with COVID-19. • Dynamic monitoring of human immune function is one of the indicators for evaluating the severity and prognosis of COVID-19 patients, Introduction Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has been widely spread. We aim to investigate the dynamic changes of peripheral blood lymphocytes subsets in adult patients with COVID-19. Methods Electronic medical records including demographics, clinical manifestation, comorbidities, laboratory data, and radiological materials of 435 hospitalized COVID-19 patients, with confirmed result of SARS-CoV-2 viral infection, were extracted and retrospectively analyzed. The counts of lymphocyte subsets at each week after the onset of the illness were compared with other weeks of illness and with those from normal individuals. Results The various lymphocyte subsets (CD3 +, CD4 +, CD8 +, CD19 +and CD16/56 +) were below the normal ranges at the first week after the onset of the illness, reaching a nadir during the second week, which increased gradually in the third week and returned to normal levels in the fifth week, but was still lower than the healthy controls. The CD3+, CD4+ and CD8+ counts were significantly lower in those with severe disease compared to those with non-severe disease, and in those patients that died compared to those who recovered. Discussion our research indicates that the levels of peripheral blood lymphocyte subsets (CD3 +, CD4 +, and CD8 +) are associated with disease progression, severity, and prognosis in patients with COVID-19. Dynamic monitoring of human immune function is one of the indicators for evaluating the severity and prognosis of COVID-19 patients, and it is helpful to formulate appropriate treatment strategies.

Purpose — to study immune status in CMV-infected pregnant women according to their detailed serologic status. Materials and methods. Serologic diagnostic method was used to determine IgM, level and avidity of IgG antibodies to CMV. ELISA was used for measurement level of TNF and TNF soluble receptors I and II. Lymphocyte subset analysis was carried out with flow cytometry. A total of 279 CMV seropositive women were examined at 6-12 weeks gestation. Results. The majority of cases (18%) of low avidity CMV IgG were found among women with positive IgM and low IgG level. The low avidity antibodies were also detected in 7% of women with positive IgM and medium IgG level, and only 1% of patients with negative IgM and high IgG level. Аvidity of IgG antibodies inversely correlated with IgM positivity (r=-0.62; p

The objective of this study was to evaluate the influence of viable cells from colostrum on immune development in dairy heifer calves during the first 28 days of life. The animals were distributed between 2 groups: COL+ (n=9) receiving fresh whole colostrum from their own damns; and COL- (n=10) receiving pooled frozen colostrum, containing no viable cells, from a pool of donor cows. These calves were assessed before colostrum intake (D0), 48 hours of age (D2), and weekly from D7 to D28. The development of immunity was evaluated by assessment of the phenotype of blood leukocyte subsets, and induced cytokine production after 72 hours of stimulation in culture with concanavalin A (ConA), killed Staphylococcus aureus (S.aureus) and killed Escherichia coli (E. coli) by peripheral blood mononuclear cell (PBMC). The clinical history of these calves was marked by a high frequency of diarrhea in both groups. However, COL- had greater diarrhea intensity scores (fecal score~3 of 4), and rectal temperature on D7 than COL+ calves. Moreover, bronchopneumonia (n=1) and navel inflammation were observed only in COL- calves. COL- had a lower concentration of serum iron, and a higher absolute number of lymphocytes on D7 than COL+. COL- also had a higher percentage of anemic calves than the COL+ calves on D21 and D28. In general, the percent of cells within each subset of leukocytes was similar between the groups over the experiment, except on week 1 when COL- calves had a higher percentage of lymphocytes expressing CD45RO+ (P=0.07). A steady increase in CD45RO+ and concomitant decline in CD45RO- leukocytes was observed over the course of the study, indicating the development of immune memory. The proportion of CD14MHCII+ leukocytes increased with age (P≤0.05). The median background cytokine production by PBMC that were not stimulated was below the level of detection of the assays used for both groups. The PBMC from COL+ calves stimulated with ConA secreted a larger quantity of IL-17 week 2 (COL+=2060.0pg/mL and COL-=0.0pg/mL, P=0.00). PBMC from COL+ calves stimulated with killed S. aureus whole cell antigen (P=0.05) and killed E. coli whole cell antigen (P=0.05) also secreted higher levels of IL17 than COL- calves at week 4. Clear production of IL17 was observed in PBML from COL+ calves at week 2, but the difference was not statistical different between groups. In conclusion, calves fed fresh and frozen colostrum showed no difference in cells subset profile overall. The increased percentage of leukocytes expressing the memory CD45RO+ or CD14MHCII+ over the course of the experiment indicated a maturation of the adaptive immune response after natural exposure to pathogens in the environment of the calf. The enhanced IL17 secretion by COL+ calves indicated that viable maternal cells modulated T-cell Th17 production that was primed by bacterial antigens. This mechanism could be responsible for quick and efficient activation of neutrophils for bacterial clearance. The differences in cytokine production observed between groups may help to explain the different clinical pictures observed for calves COL+ and COL- calves. RESUMO: O objetivo desta pesquisa foi avaliar a influência das células do colostro no desenvolvimento imune em bezerras leiteiras durante os primeiros 28 dias de vida. Os animais foram distribuídas em 2 grupos: COL+ (n=9) recebeu colostro fresco de suas próprias mães; e COL- (n=10) recebeu pool de colostro congelado sem células viáveis oriundo de vacas doadoras. Estas bezerras foram avaliadas antes da ingestão do colostro (D0); às 48 horas (D2) e semanalmente entre o D7 e D28. O desenvolvimento da imunidade foi avaliada pela fenotipagem das subpopulações celulares do sangue e produção de citocinas pelas células mononucleares sanguíneas após 72 horas de estimulação com concanavalina A (ConA), Staphylococcus aureus (S. aureus) e Escherichia coli (E. coli) inativadas. O histórico clínico das bezerras foi marcado por elevada frequência de diarreias em ambos os grupos. Entretanto, COL- apresentou maior intensidade de diarreia (escore de fezes ~ 3 de 4) e maior temperatura retal no D7 do que as bezerras COL+. Além disso, broncopneumonia (n=1) e inflamações umbilicais foram diagnosticadas apenas nas bezerras COL-. O grupo COL- apresentou menor concentração de ferro sérico e maior número de linfócitos no D7 do que o grupo COL+. COL- também apresentou maior frequência de anemias que o grupo COL+ no D21 e D28. Em geral, a fração das subpopulações celulares foram semelhantes entre os dois grupos ao longo do tempo estudado, exceto na semana 1, onde as bezerras COL- apresentaram maior proporção de CD45RO+ (P=0.07). Observou-se um constante aumento de CD45RO+ com declínio concomitante de CD45RO- ao longo do estudo, indicando o desenvolvimento da resposta imune. A proporção de células CD14MHCII+ aumentou de acordo com a idade (P≤0.05). As medianas das citocinas produzidas a partir do PBMC não estimuladas apresentaram valores abaixo do nível de detecção em ambos os grupos. O PBMC do COL+ estimulado com ConA secretou elevada concentração de IL17 na semana 2 (COL+=2060.0pg/mL e COL-=0.0pg/ml, P=0.00). PBMC do COL+ estimuladas com S. aureus inativado (P=0.05) e E. coli inativada (P=0.05) secretaram níveis mais elevados de IL17 que as bezerras COL- na semana 4. Outros picos de IL17 foram observados no COL+ na semana 2, porém não foi possível detectar diferenças estatísticas entre os grupos. Conclui-se que as bezerras alimentadas com colostro fresco e congelado apresentaram perfil similar entre as proporções das subpopulações celulares. O aumento de células expressando os marcadores de memória CD45RO+ e CD14MHCII+ demonstram, ao longo do experimento, o amadurecimento do sistema imune específico das bezerras após estimulação natural por patógenos após o nascimento. A maior secreção de IL17 pelas células das bezerras COL+ indica que as células maternas podem modular resposta imune Th17 direcionada aos antígenos bacterianos. Este mecanismo poderia ser responsável pela rápida e eficiente quimiotaxia de neutrófilos e eliminação dos microrganismos bacterianos. Os diferentes perfis de citocinas podem ser responsáveis pelos diferentes históricos clínicos relatados para as bezerras COL+ e COL-.

Introduction: Sickle cell anemia (SCA) is one of the most common genetic disorders in the world. However, until recently, only the direct consequences of the deoxyHbS polymerization was used to explain the pathophysiology of the disease. Currently, it is known that the clinical repercussions of FA involve complex interactions between the erythrocyte, endothelium and leukocytes: cytokines secreted by inflammatory cells are involved in SCA crisis and in the maintenance of a systemic inflammatory status, suggesting that T cells (Helpers and Cytotoxics) and NK have a fundalmental role in the clinical phenomena of SCA. This study aims to evaluate the profile of T and NK lymphocytes in patients with AF and compare them with the profile of individuals with sickle cell trait and individuals without hemoglobinopathies. Materials and methods: Peripheral blood was collected from 13 individuals; 7 with SS hemoglobinopathy (SS group), 5 with sickle cell trait (AS group) and 5 normal (AA group). The lymphocytes isolation for analysis was performed using Ficoll-Hypaque solution. Immunophenotyping for the determination of lymphocyte subtypes was performed by flow cytometry, using eight-color cytometer and eight BD Biosciences antibodies. Data were analyzed using Flowjo software and tabulated in SPSS IBM 22.0. The results referring to the numerical variables were expressed through measures of central tendency. Results: A lower frequency of T lymphocytes and a greater frequency of NK cells were observed in sickle cell patients.The variation of TCD4 + found between the SCA and the AA group was significant (p = 0.04). Lower frequency tendency in patients of the SS group remained for B lymphocytes. A higher frequency of NK cells was observed in patients with sickle cell anemia (mean: group AA = 15.63%, group AS = 14.82% and group SS = 23.34%) with statistically significant variation. Conclusion: SCA patients present a lower frequency of CD4 + CD8 + T cells and TNK cells when compared with HbAS individuals and individuals without hemoglobinopathies. An increasing and progressive frequency of NK cells is observed between groups AA, AS and SS. We believe that further studies are needed to understand the role of these cells in the genesis of systemic inflammation of SCA. This understanding may contribute to the development of new therapeutic strategies. INTRODUÇÃO: A anemia falciforme (AF) é uma das desordens genéticas mais comuns do mundo. Entretanto, até pouco tempo atrás, apenas as consequências diretas da polimerização da desoxiHbS explicava a fisiopatogenia da doença. Atualmente, sabe-se que as repercussões clíncas da AF envolvem interações complexas entre o eritrócito, endotélio e leucócitos: citocinas secretadas por células inflamatórias estão envolvidas nas crises e na manutenção de um status inflamatório sistêmico, o que sugere que as células T (auxiliares e citotóxicas) e NK têm papel fundalmental nos fenômenos clínicos da AF. Este estudo visa avaliar o perfil de linfócitos T e NK em portadores de AF e comparar com o perfil de indivíduos com traço falciforme e indivíduos sem hemoglobinopatias. MATERIAIS E MÉTODOS: Foi coletado sangue periférico de 17 indivíduos; 7 com hemoglobinopatia SS (grupo SS), 5 com traço falciforme (grupo AS) e 5 normais (grupo AA). Todos confirmados por eletroforese de hemoglobina. Amostras de pacientes hemotransfundidos 30 dias antes ou em uso de antiinflamatórios 2 dias antes da coleta foram excluídas. O isolamento dos linfócitos para análise foi realizado utilizando solução de Ficoll-Hypaque. A imunofenotipagem para determinação dos subtipos linfocitários foi realizada por citometria de fluxo, utilizando citômetro de oito cores e oito anticorpos da BD Biosciences. Os dados foram analisados com auxílio do software Flowjo e tabulados no SPSS IBM 22.0. Os resultados referentes às variáveis numéricas foram expressos através de medidas de tendência central: média e valores mínimos e máximos. RESULTADOS: Globalmente foi observada menor frequência de linfócitos T e maior frequência de células NK nos pacientes falcêmicos, com média de 31,2% de TCD4+ contra 43,47% do grupo AS e 45,37% do grupo AA. Para os linfócitos TCD8+, o grupo SS obteve média de 12,64% contra 17,1% do AS e 16,42% do AA. A variação de TCD4+ encontrada entre os pacientes AF e o grupo AA foi significativa (p=0,04). Tendência de menor frequência em pacientes do grupo SS se manteve para os linfócitos B. Foi observada maior frequência de células NK em pacientes portadores de anemia falciforme (média do grupo AA=8,83%; AS=11,2% e grupo SS=19,6%), respectivamente, com diferença significativa entre o grupo AA e o grupo SS (p=0,040). CONCLUSÃO: Portadores de AF apresentam tendência de menor frequência de linfócitos T, com diferença significativamente menor comparado ao portador de traço e o paciente portador de AF, para linfócitos T CD4+ verificou-se redução significativa deste subtipo celular entre grupo SS e AA. Neste estudo também foi evidenciado uma frequência crescente e progressiva de células NK entre os grupos AA, AS e SS. Acreditamos que mais estudos são necessários visando compreender o papel destas células na gênese da inflamação sistêmica da AF. Este entendimento possivelmente contribuirá para o desenvolvimento de novas estratégias terapêuticas. Aracaju, SE

Purpose — to study immune status in CMV-infected pregnant women according to their detailed serologic status.Materials and methods. Serologic diagnostic method was used to determine IgM, level and avidity of IgG antibodies to CMV. ELISA was used for measurement level of TNF and TNF soluble receptors I and II. Lymphocyte subset analysis was carried out with flow cytometry. A total of 279 CMV seropositive women were examined at 6-12 weeks gestation.Results. The majority of cases (18%) of low avidity CMV IgG were found among women with positive IgM and low IgG level. The low avidity antibodies were also detected in 7% of women with positive IgM and medium IgG level, and only 1% of patients with negative IgM and high IgG level. Аvidity of IgG antibodies inversely correlated with IgM positivity (r=-0.62; p, Цель — изучить состояние иммунной системы беременных женщин в зависимости от их детализированного ЦМВ-серологичеcкого статуса.Пациенты и методы. Метод серологической диагностики использован для определения наличия IgM, уровня и авидности IgG антител к ЦМВ. Иммуноферментный анализ использован для измерения уровня ФНО и его растворимых рецепторов I и II типа. Анализ субпопуляций лимфоцитов осуществлен проточной цитометрией. В общей сложности 279 ЦМВ-серопозитивных женщин обследованы в сроке 6–12 недель беременности.Результаты. Большинство (18%) случаев низкой авидности ЦМВ IgG обнаружены среди женщин с наличием IgM и низким уровнем IgG. У 7% женщин с наличием IgM и средним уровнем IgG и только у 1% женщин с отсутствием IgM и высоким уровнем IgG отмечены низкоавидные антитела. Авидность IgG антител обратно коррелировала с IgM позитивностью (r=-0,62; р, Мета — вивчити стан імунної системи у вагітних жінок залежно від їхнього деталізованого ЦМВ-серологічного статусу.Пацієнти та методи. Метод серологічної діагностики використано для визначення наявності IgM, рівня та авідності IgG антитіл до ЦМВ. Імуноферментний аналіз використано для вимірювання рівня ФНП і його розчинних рецепторів I і II типу. Аналіз субпопуляцій лімфоцитів здійснено проточною цитометрією. У цілому 279 ЦМВ-серопозитивних жінок обстежено в терміні 6–12 тижнів вагітності.Результати. Більшість (18%) випадків низької авідності ЦМВ IgG виявлено серед жінок із наявністю IgM і низьким рівнем IgG. У 7% жінок із наявним IgM і середнім рівнем IgG та лише в 1% жінок із відсутнім IgM і високим рівнем IgG також визначено низькоавідні антитіла. Авідність IgG антитіл зворотно корелювала з IgM позитивністю (r=-0,62; р

The objective of this study was to evaluate the influence of viable cells from colostrum on immune development in dairy heifer calves during the first 28 days of life. The animals were distributed between 2 groups: COL+ (n=9) receiving fresh whole colostrum from their own damns; and COL- (n=10) receiving pooled frozen colostrum, containing no viable cells, from a pool of donor cows. These calves were assessed before colostrum intake (D0), 48 hours of age (D2), and weekly from D7 to D28. The development of immunity was evaluated by assessment of the phenotype of blood leukocyte subsets, and induced cytokine production after 72 hours of stimulation in culture with concanavalin A (ConA), killed Staphylococcus aureus (S.aureus) and killed Escherichia coli (E. coli) by peripheral blood mononuclear cell (PBMC). The clinical history of these calves was marked by a high frequency of diarrhea in both groups. However, COL- had greater diarrhea intensity scores (fecal score~3 of 4), and rectal temperature on D7 than COL+ calves. Moreover, bronchopneumonia (n=1) and navel inflammation were observed only in COL- calves. COL- had a lower concentration of serum iron, and a higher absolute number of lymphocytes on D7 than COL+. COL- also had a higher percentage of anemic calves than the COL+ calves on D21 and D28. In general, the percent of cells within each subset of leukocytes was similar between the groups over the experiment, except on week 1 when COL- calves had a higher percentage of lymphocytes expressing CD45RO+ (P=0.07). A steady increase in CD45RO+ and concomitant decline in CD45RO- leukocytes was observed over the course of the study, indicating the development of immune memory. The proportion of CD14MHCII+ leukocytes increased with age (P≤0.05). The median background cytokine production by PBMC that were not stimulated was below the level of detection of the assays used for both groups. The PBMC from COL+ calves stimulated with ConA secreted a larger quantity of IL-17 week 2 (COL+=2060.0pg/mL and COL-=0.0pg/mL, P=0.00). PBMC from COL+ calves stimulated with killed S. aureus whole cell antigen (P=0.05) and killed E. coli whole cell antigen (P=0.05) also secreted higher levels of IL17 than COL- calves at week 4. Clear production of IL17 was observed in PBML from COL+ calves at week 2, but the difference was not statistical different between groups. In conclusion, calves fed fresh and frozen colostrum showed no difference in cells subset profile overall. The increased percentage of leukocytes expressing the memory CD45RO+ or CD14MHCII+ over the course of the experiment indicated a maturation of the adaptive immune response after natural exposure to pathogens in the environment of the calf. The enhanced IL17 secretion by COL+ calves indicated that viable maternal cells modulated T-cell Th17 production that was primed by bacterial antigens. This mechanism could be responsible for quick and efficient activation of neutrophils for bacterial clearance. The differences in cytokine production observed between groups may help to explain the different clinical pictures observed for calves COL+ and COL- calves.

Background: The importance of lymphocyte subtypes in determining outcome in primary operable ductal invasive breast cancer remains unclear. The aim of present study was to examine the relationship between tumour lymphocyte subsets infiltrate and standard clinico-pathological factors and survival in patients with primary operable invasive ductal breast cancer. Methods: The analysis of the inflammatory cell infiltrate, including lymphocyte subtypes, was undertaken using immunohistochemical techniques and visual quantitative and semi-quantitative techniques in 338 patients with ductal breast cancer. Results: The majority (91%) of patients had high grade inflammatory cell infiltrate. The median follow-up of the survivors was 164 months. During this period, 65 died of their cancer. On univariate analysis, tumour inflammatory cell infiltrate, macrophages infiltrate (P

Background: Host effector mechanism against Mycobacterium tuberculosis (Mtb) infection is dependent on innate immune response by macrophages and neutrophils and the alterations in balanced adaptive immunity. Coordinated release of cytolytic effector molecules from NK cells and effector T cells and the subsequent granule-associated killing of infected cells have been documented; however, their role in clinical tuberculosis (TB) is still controversy. Objective: To investigate whether circulating granulysin and other effector molecules are associated with the number of NK cells, iNKT cells, Vγ9+Vδ2+ T cells, CD4+ T cells and CD8+ T cells, and such association influences the clinical outcome of the disease in patients with pulmonary TB and HIV/TB coinfection. Methods: Circulating granulysin, perforin, granzyme-B and IFN-γ levels were determined by ELISA. The isoforms of granulysin were analyzed by Western blot analysis. The effector cells were analyzed by flow cytometry. Results: Circulating granulysin and perforin levels in TB patients were lower than healthy controls, whereas the granulysin levels in HIV/TB coinfection were much higher than in any other groups, TB and HIV with or without receiving HAART, which corresponded to the number of CD8+ T cells which kept high, but not with NK cells and other possible cellular sources of granulysin. In addition, the 17kDa, 15kDa and 9kDa isoforms of granulysin were recognized in plasma of HIV/TB coinfection. Increased granulysin and decreased IFN-γ levels in HIV/TB coinfection and TB after completion of anti-TB therapy were observed. Conclusion: The results suggested that the alteration of circulating granulysin has potential function in host immune response against TB and HIV/TB coinfection. This is the first demonstration so far of granulysin in HIV/TB coinfection.

Understanding the impact of training sessions on the immune response is crucial for the adequate periodization of training, to prevent both a negative influence on health and a performance impairment of the athlete. This study evaluated acute systemic immune cell changes in response to an actual swimming session, during a 24-h recovery period, controlling for sex, menstrual cycle phases, maturity, and age group. Competitive swimmers (30 females, 15 ± 1.3 years old; and 35 males, 16.5 ± 2.1 years old) performed a high-intensity training session. Blood samples were collected before, immediately after, 2 h after, and 24 h after exercise. Standard procedures for the assessment of leukogram by automated counting (Coulter LH 750, Beckman) and lymphocytes subsets by flow cytometry (FACS Calibur BD, Biosciences) were used. Subjects were grouped according to competitive age groups and pubertal Tanner stages. Menstrual cycle phase was monitored. The training session induced neutrophilia, lymphopenia, and a low eosinophil count, lasting for at least 2 h, independent of sex and maturity. At 24 h postexercise, the acquired immunity of juniors (15–17 years old), expressed by total lymphocytes and total T lymphocytes (CD3+), was not fully recovered. This should be accounted for when planning a weekly training program. The observed lymphopenia suggests a lower immune surveillance at the end of the session that may depress the immunity of athletes, highlighting the need for extra care when athletes are exposed to aggressive environmental agents such as swimming pools.

Environmental studies performed in Panasqueira mine area (central Portugal) identified high concentrations of several metal(loid)s in environmental media, and individuals environmentally and occupationally exposed showed higher levels of As, Cr, Mg, Mn, Mo, Pb and Zn in blood, urine, hair and nails when compared to unexposed controls. To evaluate the presence of immunological alterations attributable to environmental contamination, we quantified neopterin, kynurenine, tryptophan, and nitrite concentrations in plasma, and analysed the percentage of several lymphocytes subsets, namely CD3+, CD4+ and CD8+ T-cells, CD19+ B-cells, and CD16+56+ natural killer (NK) cells in a group of individuals previously tested for metal(loid) levels in different biological matrices. The environmentally exposed group had significantly lower levels of %CD8+ and higher CD4+/CD8+ ratios, whereas the occupationally exposed individuals showed significant decreases in %CD3+ and %CD4+, and significant increases in %CD16+56+, when compared to controls. Analysed biomarkers were found to be influenced by age, particularly neopterin, kynurenine and kynurenine to tryptophan ratio (Kyn/Trp) with significantly higher levels in older individuals, and %CD3+, %CD8+ and %CD19+ with significantly lower values in older individuals. Males environmentally exposed showed significantly lower values of %CD19+ when compared to control females. The concentration of Pb in toenails was associated to the level of neopterin, kynurenine and Kyn/Trp ratio (all direct), and the concentration of Mn in blood to the level of %CD8+, %CD19+ (both inverse) and CD4+/CD8+ ratio (direct). Overall our results show that the metal(loid) contamination in Panasqueira mine area induced immunotoxic effects in exposed populations, possibly increasing susceptibility to diseases. This work was supported by the Portuguese Foundation for Science and Technology under the grant SFRH/BD/47781/2008 and the project PTDC/SAU-ESA/102367/2008, and by the European Commission (ERA NET — New INDIGO Program, NanoLINEN project, PIM2010ENI-00632). The work of Roberto Zoffoli, Valentina Dall'Armi, and Stefano Bonassi was supported by grants funded by the Associazione Italiana per la Ricerca sul Cancro (AIRC).

OBJECTIVE To investigate the lymphocyte subpopulations (T4, T8 and macrophages) and major histocompatibility (MHC) II antigens in patients with superficial bladder cancer before and after intravesical instillations of recombinant interferon-gamma (IFN-gamma). PATIENTS AND METHODS Four intravesical weekly instillations of either 1.3 mg (20 patients, group A) or 0.7 mg (11 patients, group B) IFN-gamma were administered in 31 evaluable patients (28 men and three women, mean age 68.5 years). The CD4+, CD8+, CD68+ and HLA-DR antigens were detected immunohistochemically in tumours and a marker tumour before and after intravesical instillations. RESULTS The median number of T4 lymphocytes increased from 15 per high-power field (HPF) to 27.5 in group A (P = 0.0029) and to 45 in group B (P = 0.0117). Macrophages increased from 6 cells/HPF to 15 cells/HPF in group A (P = 0.0029) and from 2 to 8.75 cells/HPF in group B (P = 0.0117). The T8 lymphocyte subpopulation decreased from 4 to 3 cells/HPF (P = 0.0231) in group A and from 5 to 2 cells/HPF (P = 0.0759) in group B. The median percentage of HLA-DR antigens increased from 1.5% to 18% in general, (P < 0.001), from 2.5% to 15% in group A (P = 0.0064) and from 0% to 20% in group B (P = 0.0077). The induction of HLA-DR antigens was statistically significant in those receiving the lower dose (from 0% before instillation to 20% afterward, P = 0.0277), while it was not with the higher dose (from 0% to 5%, P = 0.068). Irrespective of the dose of IFN used. T4 lymphocytes and macrophages increased significantly after treatment in patients in whom the tumour HLA-DR antigens were either up-regulated or remained stable. The median net increase in T4 cells was 17.5 and 30 cells/HPF for groups A and B, respectively (P = 0.0429). CONCLUSION T4 lymphocytes, macrophages and HLA-DR antigens increased after intravesical IFN-gamma in patients with superficial bladder cancer, but T8 lymphocytes decreased. Irrespective of the drug dose used, patients with either upregulated or stable HLA-DR antigens after treatment showed the same pattern of changes in the lymphocyte subpopulations. The two doses generally had the same effect on the immunological variables assessed but the lower dose was more effective in inducing HLA-DR antigens and in increasing the number of T4 lymphocytes in the tumours.

Ο σκοπός αυτής της μελέτης ήταν να διερευνηθεί η συχνότητα της εμφάνισης λοίμωξης σε ηπατοπαθείς, η καταγραφή των πιο συχνών λοιμώξεων σε αυτή την ομάδα ασθενών καθώς και η διερεύνηση της επίπτωσης της κίρρωσης στην κυτταρική τους ανοσία Επίσης, στόχος μας ήταν η διερεύνηση της παρουσίας των κυτοκινών στο ασκιτικό υγρό κιρρωτικών ασθενών με ή χωρίς αυτόματη βακτηριακή περιτονίτιδα (ΑΒΠ) και της επίδρασης της ΑΒΠ στην έκφραση των προφλεγμονωδών, αντιφλεγμονωδών κυτοκινών και των υποδοχέων τους στο ασκιτικό υγρό ΑΣΘΕΝΕΙΣ-ΜΕΘΟΔΟΙ: Η μελέτη περιέλαβε ασθενείς με κίρρωση του ήπατος, οποιασδήποτε αιτιολογίας, που εισήχθησαν στο νοσοκομείο από το Μάιο 2006 μέχρι τον Δεκέμβριο 2007. Από τους συνολικά 110 ασθενείς με κίρρωση, οι 67 (ομάδα 1, 59.1%) εισήχθησαν για διάφορους λόγους εκτός από λοίμωξη, οι 24 (ομάδα 2, 21.8%) παρουσίασαν σύνδρομο σήψης και οι 19 (ομάδα 3, 19.1%) παρουσίασαν κιρσορραγία. Σε όλους τους ασθενείς έγινε καταγραφή των δημογραφικών τους στοιχείων, του τύπου λοίμωξης και μετρήθηκαν οι Τ- λεμφοκυτταρικοί υποπληθυσμοί, δηλ. CD3, CD4, CD5, CD8, CD14, CD20, CD56, CD64 καθώς και ο λόγος CD4/CD8 με κυτταρομετρία ροής. Οι μετρήσεις στους ασθενείς των ομάδων 2 και 3 έγιναν κατά την ημέρα της εισαγωγής, την τρίτη ημέρα νοσηλείας και την ημέρα εξόδου. Στους ασθενείς της ομάδας 1, οι μετρήσεις έγιναν μόνο την ημέρα της εισαγωγής. Μελετήθηκαν επίσης και 30 υγιή άτομα της ίδιας εθνικότητας και με παρόμοια δημογραφικά χαρακτηριστικά με τους υπόλοιπους ασθενείς. Αυτοί δεν νοσηλέυθηκαν, δεν είχαν κανένα πρόβλημα υγείας ούτε είχαν μεταγγισθεί. Μελετήθηκαν επίσης χωριστά 13 κιρρωτικοί ασθενείς, από τους συνολικά 110, οι οποίοι είχαν ασκίτη και εισήχθησαν στο Νοσοκομείο κατά την ίδια περίοδο. Αυτοί διακρίθηκαν σε δυο ομάδες: ομάδα 1 : ασθενείς με ΑΒΠ , ομάδα 2: ασθενείς χωρίς ΑΒΠ. Μετρήθηκαν οι κυττοκίνες : IL-1b, IL-1RA, IL-6, IL-10, TGF-1b, TNF-a, STNFRI, STNFRII. Οι μετρήσεις έγιναν με μέθοδο ELISA. ΑΠΟΤΕΛΕΣΜΑΤΑ : Οι απόλυτες τιμές των ολικών Τ λεμφοκυττάρων (CD3), των βοηθητικών (CD4) λεμφοκυττάρων, των κατασταλτικών (CD8) λεμφοκυττάρων και των CD5 λεμφοκυττάρων είναι μικρότερες στις ομάδες των κιρρωτικών με λοίμωξη και κιρσορραγία. Ειδικότερα, στην ομάδα των κιρσορραγούντων κιρρωτικών οι διαφορές στα βοηθητικά λεμφοκύτταρα (CD4) και τα ολικά λεμφοκύτταρα (CD3) κατά την έξοδο από το νοσοκομείο ήταν στατιστικά σημαντικές (Ρ< 0,05), ενώ όχι στους σηπτικούς. Οι απόλυτες τιμές όλων των Τ λεμφοκυτταρικών υποπληθυσμών ήταν υψηλότερες κατά την εισαγωγή και μειώθηκαν κατά τη διάρκεια νοσηλείας. Οι διαφορές στο ποσοστό συνέκφρασης στους δείκτες CD14/CD64 των ουδετεροφίλων ήταν σημαντικά μειωμένο στους σηπτικούς.Οι τιμές των κυτοκινών IL-10, IL-6, TNF-a, IL-1RA, STNFRI, STNFRII ήταν υψηλότερες στους ασθενείς με ΑΒΠ. Στατιστικά σημαντικές διαφορές (Ρ

Probiotic research has been approached, above all in recent years, by widely differing points of view, both for human and animal uses. Lactic acid bacteria release bacteriocins, and some of them may function as probiotic. The aim of the present study was to investigate the effects of dietary supplementation with the probiotic Pediococcus acidilactici on the piglet intestine, on circulating lymphocytes, and on aspects of piglet performance during the first 42 days after weaning. Sixteen female piglets were at weaning assigned to two dietary groups: Control (Ctr, 8 animals) and Pediococcus acidilactici supplemented (Pa, 8 animals). Piglets’ growth was monitored from weaning to the end of the trial. On day 42 post-weaning, the piglets were slaughtered and small specimens from both ileum and cecum were examined with haematoxylin/eosin staining to ascertain structural details. Histometry was performed by villi and crypts measurements, as well as GALT measurements. Histochemical analyses were performed to investigate the intestinal mucins. Immunohistochemical analyses were used to visualize proliferating as well as apoptotic mucosal cells, and to identify mucosal macrophages and IgA producing cells. Intra-epithelial CD8+ T lymphocytes were identified and counted. Subsets of circulating T lymphocytes were analyzed by flow cytometry. Pediococcus acidilactici supplementation positively influenced weight and post-weaning average daily gain of treated piglets. Histometry showed an increase in villi height and crypts depth in Pa animals in comparison with controls. Treated piglets showed a larger number of proliferating enterocytes than controls. Intra-epithelial CD8+ T lymphocytes were scarcer in treated than in control piglets, likely in relation with catarrhal enteritis shown in the latter. We conclude that the studied supplementation was possibly able to protect the piglet small intestinal mucosa, improving local resistance to infections in the stressful weaning period.

Metallothionein (MT) expression in intestinal resection specimens from 41 patients with ulcerative colitis (UC) and 10 patients with Crohn’s disease (CD ) was immunohistochemically studied by the avidin-biotin (ABC) method. In addition, the possible relationship of its expression with HLA-DR antigen expression, lymphocyte subpopulations and proliferation-associated indices was studied in order to elucidate the role of this molecule in inflammatory bowel disease (IBD). The MT immunoreactivity was recorded by staining and intensity-distribution scores. MT staining varied in and was mainly localized in the cytoplasm, although a combined nuclear/cytoplasmic reactive pattern was also seen in epithelial cells. MT expression was decreased in UC, and CD compared with normal mucosa. No difference in MT expression between UC and CD was noted. In UC, a gradually decreased expression from remission, to resolving and to active phase was observed. An inverse correlation of MT expression with HLA-DR antigen expression was detected (p=0.018) in the cases of UC. The data suggest that a low level of MT expression in inflammatory bowel disease and particularly in active phase of UC may indicate a decreased endogenous intestinal protection and it may be implicated in the pathogenesis of the disease.

RESUMO Esta pesquisa avaliou a dinâmica dos leucócitos e das subpopulações de linfócitos em vacas Holandesas soropositivas para o BLV no período de transição. Amostras de sangue (n=72) provenientes de 12 vacas foram coletadas entre as semanas -2 e +3 para a realização do leucograma, imunofenotipagem, dosagem de cortisol e haptoglobina (Hp). O perfil leucocitário foi caracterizado por leucocitose, neutrofilia, monocitose e eosinopenia próximo ao parto. Linfocitose e elevada proporção de linfócitos B CD21+ foram achados constantes entre as semanas -2 e +3; assim, as vacas foram testadas e confirmadas soropositivas para o BLV. Os valores das subpopulações de linfócitos T apresentaram-se baixos durante o período de transição, observando-se dois picos máximos que coincidiram com as elevações nas concentrações de cortisol no parto (2,11µg/dL) e semana +3 (1,97µg/dL). Hp apresentou aumento crescente de -2 (166µg/mL) a +3 (576µg/mL), provavelmente associada à elevada taxa de infecções uterinas observadas nas semanas +2 e +3. As vacas soropositivas para o BLV apresentaram leucograma de estresse próximo ao parto, exceto para linfócitos. A linfocitose e as elevadas proporções de células B CD21+, associadas com as baixas proporções de células T, podem ser indicativo de imunossupressão e predisposição aos processos inflamatórios no período pós-parto. ABSTRACT This research evaluated the dynamics of leukocytes and lymphocytes subsets in seropositive Holstein cows for BLV during the transition period. Blood samples (n=72) from 12 cows were harvested from week -2 up to week +3 to perform leukogram, immunophenotyping, cortisol and haptoglobin (Hp). Leukocytes pattern was characterized by leukocytosis, neutrophilia, monocytosis and eosinopenia around calving. Lymphocytosis and high proportions of B cells CD21+ were a constant finding between week -2 and +3, thus cows were tested and confirmed seropositive for BLV. The values of T lymphocytes subsets were low during the transition period, observing two peaks that coincided with high levels of cortisol at delivery (2.11µg/dL) and week +3 (1.97µg/dL). Hp had gradual increase from week -2 (166µg/mL) until week +3 (576g/mL) probably due to high rate of uterine infection detected between week +2 and +3. The seropositive cows for BLV presented stress leukogram around delivery, except for lymphocytes. Lymphocytosis and the high proportions of B cells, associated with the low proportions of T lymphocytes, can be indicative of immunosuppression and predisposition to the inflammatory process observed in the post-partum period.