Imuninė sensitizacija po dirbtinės širdies prijungimo

Radvilė Malickaitė1,2, Aldona Stanevičienė2, Kęstutis Ručinskas1,2, Laimutė Jurgauskienė1,2, Saulius Miniauskas1,2, Vytautas Sirvydis1,2 1 Vilniaus universiteto Širdies ir kraujagyslių ligų klinikos Širdies chirurgijos centras, Santariškių g. 2, LT-08661 Vilnius 2 Vilniaus universiteto ligoninės Santariškių klinikos, Santariškių g. 2, LT-08661 Vilnius El paštas: [email protected] Tikslas Įvertinti Vilniaus universiteto ligoninės Santariškių klinikų Širdies chirurgijos centre atliekamos dirbtinės širdies implantavimo procedūros įtaką imuninei širdies laukiančių recipientų būklei ir palyginti gautus rezultatus su kitų centrų duomenimis. Ligoniai ir metodai Ištirta aštuonių potencialių širdies recipientų, kurių kraujotaka buvo palaikoma dirbtinės širdies skilvelio (DŠS), imuninių rodiklių dinamika: prieš DŠS implantavimą ir praėjus ne mažiau kaip vienam mėnesiui po operacijos. Nustatytas periferinio kraujo imunokompetentinių ląstelių CD3+CD4+ ir CD3+CD8+ skaičius, Apo-1/FAS (CD95) ekspresija CD4+ ir CD8+ limfocitų paviršiuje, viduląstelinė aktyvi kaspazė-3 ir sensitizacija HLA antigenams. Rezultatai Po DŠS prijungimo atsiranda limfocitų T helperių (CD3+CD4+) skaičiaus trūkumas, ryškesnis pacientų, kuriems implantuotas išorinis dirbtinis skilvelis. Numanomas ląstelių praradimo mechanizmas yra apoptozės indukcija. Nepaisant ilgalaikio DŠS prijungimo ir pakartotinių kraujo produktų perpylimų (lentelė), antikūnai prieš HLA sistemos antigenus daugumai pacientų nesigamino: sensitizacija nustatyta tik dviem iš aštuonių pacientų (gavusių santykiškai daugiau kraujo produktų). Išvada Sensitizacija apsunkina pacientų gydymą, ypač vengtini kraujo produktai. Patartina naudoti leukofiltruotus kraujo produktus. Pagrindiniai žodžiai: dirbtiniai širdies skilveliai, imunologinė sensitizacija, apoptozė Comparison of quality of life and physical condition of heart failure Radvilė Malickaitė1,2, Aldona Stanevičienė2, Kęstutis Ručinskas1,2, Laimutė Jurgauskienė1,2, Saulius Miniauskas1,2, Vytautas Sirvydis1,2 1 Vilnius University Clinic of Cardiovascular Diseases, Cardiac Surgery Centre, Santariškių str. 2, LT-08661 Vilnius, Lithuania 2 Vilnius University Hospital „Santariškių klinikos“, Santariškių str. 2, LT-08661 Vilnius, Lithuania E-mail: [email protected] Objective This study was designed to characterize alterations in the immune system of patients who received artificial heart (AH) placement therapy for end-stage heart failure as a bridge to heart transplantation at the Vilnius University Cardial Surgery Centre. Patients and methods Immunological studies were performed in 8 pts awaiting cardiac transplantation with AH assist (7 of them were transplanted after 75–1050 days of support). Immunological indices before AH placement and after one month of mechanical assistance (period free from infection) were compared. Fluorochrome-labeled Mabs were used for immunophenotypic analysis of circulating T cells and apoptocic activity evaluation. Antibodies against HLA class I molecules were detected by a standard microlymphocytotoxic test. Results AH recipients showed relative lymphopenia, reduction in CD4 T cell counts, a decreased CD+4+ / CD8+ ratio. Six of the eight recipients with long-term support did not produce HLA antibodies despite receiving blood products during support. Conclusions In conclusion, these results suggest that reduction in CD4+ T cell levels accompanying AH support may increase the prevalence of infectious complications and HLA sensitization. Key words: heart-assist devices, immune, sensitization, apoptosi

Imuninė stebėsena širdies transplantacijoje: atmetimo reakcijos poveikis T limfocitų žymenų ekspresijai

Radvilė Malickaitė, Laimutė Jurgauskienė, Stanislava Simanavičienė, Vytė Valerija Maneikienė, Rita Sudikienė, Kęstutis Ručinskas Vilniaus universiteto Širdies ir kraujagyslių ligų klinikos Širdies chirurgijos centras, Santariškių g. 2, LT-08661 Vilnius El. paštas: [email protected] Darbo tikslas: Nustatyti Vilniaus universiteto ligoninės Santariškių klinikų Širdies chirurgijos centre atliekamos širdies transplantacijos įtaką T limfocitų aktyvumo rodikliams, įvertinti imuninės stebėsenos tinkamumą ūminiam transplantato atmetimui prognozuoti. Ligoniai ir metodai: Retrospektyviai analizuotas dvidešimt vieno širdies recipiento imuninių rodiklių kitimas esant normaliai potransplantacinei būklei ir ūminiam transplantato atmetimui. Periferinio kraujo imunokompetentinių ląstelių CD3+CD103+, CD4+CD103+, CD8+CD103+, CD3+CD134+, CD4+CD134+, CD8+CD134+, CD8+CD57+ ir CD8+CD38+ procentas nustatytas tėkmės citometrijos būdu. Ūminis transplantato atmetimas vertintas pagal histologinius endomiokardinės biopsijos radinius. Rezultatai: Esant ūminio atmetimo epizodams, kai endomiokardo biopsijos įvertintos ≥ 2R (3A) laipsniu, reikšmingai didėja integrino CD103 (p < 0,0001), kostimuliacinio receptoriaus CD134 (p = 0,005), antigeno CD57 (p = 0,005) ir ląstelių paviršiaus glikoproteino CD38 (p = 0,015) ekspresija citotoksinių CD8+ limfocitų paviršiuje. Išvados: Imuninė periferinio kraujo limfocitų būklės stebėsena gali būti taikoma po transplantacijos skiriamam imunosupresiniam gydymui įvertinti numatant didelę ūminio atmetimo tikimybę. Reikšminiai žodžiai: kiaušidžių cistos, supiktybėjimo rizika, piktybiškumo rizikos indeksas, ultragarsinis tyrimas, Ca-125 antigenas, chirurginis gydymas Measuring T cell reactivity for predicting heart transplant rejection Radvilė Malickaitė, Laimutė Jurgauskienė, Stanislava Simanavičienė, Vytė Valerija Maneikienė, Rita Sudikienė, Kęstutis Ručinskas Vilnius University, Clinic of Cardiovascular Diseases, Centre of Heart sSurgery, Santariškių str. 2, LT-08661 Vilnius, Lithuania E-mail: [email protected] Objective: We aimed to analyze alterations in peripheral blood T-cell subset activation compared with endomyocardial byopsy findings. Patients and methods: The study included in total twenty-one heart recipients grafted 1997–2007 at the Vilnius Heart sSurgery cCenter. T-cell activation markers CD3+CD103+, CD4+CD103+, CD8+CD103+, CD3+CD134+, CD4+CD134+, CD8+CD134+, CD8+CD57+ ir CD8+CD38+ were detected by two-color flow cytometry. Rejection was graded according to the ISHLT (the International Society of Heart and Lung Transplantation) grading system. Results: In case of ≥ 2R (3A) rejection episodes, a significant increase in the expression of integrin CD103 (p < 0.0001), co-stimulatory receptor CD134 (p = 0.005), antigen CD57 (p = 0.005) and surface glycoprotein CD38 (p = 0.015) on CD8+ T lymphocytes has been revealed. Conclusion: Immune monitoring performed on peripheral blood can be used for the assessment of immunosuppression therapy on transplant recipients’ immune response and for determining the risk of rejection. Key words: heart transplantation, acute rejection, and immune activatio

Chronic urticaria and thyroid autoimmunity markers

The association of chronic urticaria and autoimmune thyroid disease has been well recognized. Some authors observed relationship between thyroid autoimmunity and chronic autoimmune urticaria

Imunomitybos poveikis T ląstelių aktyvacijai: atsitiktinių imčių tyrimas kardiochirurginių ligonių grupėje

Background. Cardiac surgery provokes an intense inflammatory response that can cause an immunosuppressive state and adverse postoperative outcomes. We recently showed that postoperative immunonutrition with glutamine in “fragile” low-risk cardiac surgery patients was associated with a significantly increased level of CD3+ and CD4+ T cells. In order to clarify the biological relevance and clinical importance of these findings, we investigated whether an increase in the CD4+ T cell level was caused by changes in the systemic inflammatory response (caused by surgery or infection) and if it was associated with their activation status. Methods. A randomized control study of low operative risk but “fragile” cardiac surgery patients was performed. Patients were randomized into immunonutrition (IN) and control groups (C). The IN group received normal daily meals plus special immune nutrients for 5 days postoperatively, while the C group received only normal daily meals. Laboratory parameters were investigated before surgery and on the sixth postoperative day and the groups were compared accordingly. The expression of the CD69+ marker was investigated to determine T cell activation status. Serum concentrations of cytokines (interleukin-10 (IL-10), tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6)) and C-reactive protein (CRP) were determined to assess the systemic inflammatory response, while procalcitonin (PCT) levels were evaluated to confirm or deny possible bacterial infection. Results. Fifty-five patients were enrolled in the study. Twenty-seven (49.1%) were randomized in the IN group. Results show that on the sixth postoperative day, the CD4+CD69+ and CD8+CD69+ counts did not differ between the IN and C groups, accordingly 0.25 [0.16–0.50] vs 0.22 [0.13-0.41], p=0.578 and 0.13 [0.06–0.3] vs 0.09 [0.05–0.14], p=0.178. Also, statistically significant differences were not observed in the cytokine levels (IN and C groups: TNF-α 8.13 [7.32–10.31] vs 8.78 [7.65–11.2], p=0.300; IL-6 14.65 [9.28–18.95] vs 12.25 [8.55–22.50], p=0.786; IL-10 5.0 [5.0–5.0] vs 5.0 [5.0–5.0], p=0.343 respectively), which imply that an elevated T cell count is not associated with the systemic inflammatory response. Also, PCT (IN and C groups: 0.03 [0.01–0.09] vs 0.05 [0.03–0.08], p=0.352) and CRP (IN and C groups 62.7 [34.2–106.0] vs 63.7 [32.9–91.0], p=0.840) levels did not differ between the two groups. Moreover, low levels of PCT indicated that the increase in T cell count was not determined by bacterial infection. Conclusions. Our findings showed that CD4+ T cell levels were associated with neither the systemic inflammatory response nor bacterial infection. Secondly, increases in T cells are not accompanied by their activation status. These results suggest a hypothesis that a higher postoperative T cell concentration may be associated with postoperative immunonutrition in low-risk cardiac surgery patients with intact cellular vitality, i.e. “fragile”. However, immunonutrition alone did not affect T cell activation status

CD31+, CD38+, CD44+, and CD103+ lymphocytes in peripheral blood, bronchoalveolar lavage fluid and lung biopsy tissue in sarcoid patients and controls

Background: The mechanisms driving the transition from inflammation to fibrosis in sarcoidosis patients are poorly understood; prognostic features are lacking. Immune cell profiling may provide insights into pathogenesis and prognostic factors of the disease. This study aimed to establish associations in simultaneous of lymphocyte subset profiles in the blood, bronchoalveolar lavage fluid (BALF), and lung biopsy tissue in the patients with newly diagnosed sarcoidosis. Methods: A total of 71 sarcoid patients (SPs) and 20 healthy controls (HCs) were enrolled into the study. CD31, CD38, CD44, CD103 positive T lymphocytes in blood and BALF were analysed. Additionally, the densities of CD4, CD8, CD38, CD44, CD103 positive cells in lung tissue biopsies were estimated by digital image analysis. Results: Main findings: (I) increase of percentage of CD3+CD4+CD38+ in BALF and blood, and increase of percentage of CD3+CD4+CD44+ in BALF in Löfgren syndrome patients comparing with patients without Löfgren syndrome, (II) increase of percentage of CD3+CD4+103+ in BALF and in blood in patients without Löfgren syndrome (comparing with Löfgren syndrome patients) and increase of percentage of CD3+CD4+103+ in BALF and in blood in more advanced sarcoidosis stage. (III) Increasing percentage of BALF CD3+CD4+CD31+ in sarcoidosis patients when comparing with controls independently of presence of Löfgren syndrome, smoking status or stage of sarcoidosis. Several significant correlations were found. Conclusions: Lymphocyte subpopulations in blood, BALF, and lung tissue were substantially different in SPs at the time of diagnosis compared to HCs. CD3+CD4+CD31+ in BALF might be a potential supporting marker for the diagnosis of sarcoidosis. CD3+CD4+CD38+ in BALF and blood and CD3+CD4+CD44+ in BALF may be markers of the acute immune response in sarcoidosis patients. CD4+CD103+ T-cells in BALF and in blood are markers of the persistent immune response in sarcoidosis patients and are potential prognostic features of the chronic course of this disease

Lymphocyte subsets and pulmonary nodules to predict the progression of sarcoidosis

The search for biological markers, which allow a relatively accurate assessment of the individual course of pulmonary sarcoidosis at the time of diagnosis, remains one of the research priorities in this field of pulmonary medicine. The aim of our study was to investigate possible prognostic factors for pulmonary sarcoidosis with a special focus on cellular immune inflammation markers. A 2-year follow-up of the study population after the initial prospective and simultaneous analysis of lymphocyte activation markers expression in the blood, as well as bronchoalveolar lavage fluid (BALF) and lung biopsy tissue of patients with newly diagnosed pulmonary sarcoidosis, was performed. We found that some blood and BAL fluid immunological markers and lung computed tomography (CT) patterns have been associated with a different course of sarcoidosis. We revealed five markers that had a significant negative association with the course of sarcoidosis (worsening pulmonary function tests and/or the chest CT changes)—blood CD4+CD31+ and CD4+CD44+ T lymphocytes, BALF CD8+CD31+ and CD8+CD103+ T lymphocytes and a number of lung nodules on chest CT at the time of the diagnosis. Cut-off values, sensitivity, specificity and odds ratio for predictors of sarcoidosis progression were calculated. These markers may be reasonable predictors of sarcoidosis progression