| 1. |
- Kvarnström, Maria, 1971-, et al.
(författare)
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Effect of cryopreservation on expression of Th1 and Th2 cytokines in blood mononuclear cells from patients with different cytokine profiles, analysed with three common assays: an overall decrease of interleukin-4 : An overall decrease of interleukin-4
- 2004
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Ingår i: Cryobiology. - : Elsevier BV. - 0011-2240 .- 1090-2392. ; 49:2, s. 157-168
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Tidskriftsartikel (refereegranskat)abstract
- Studies on cytokine expression in blood cells are commonly performed on cryopreserved cells. Previous studies show that cryopreservation affects cytokine expression, but the findings are not consistent. This may be due to divergent effects of freezing on different cytokines, different stimuli, and different patient groups or to the use of different assays in the studies. This study was designed to investigate the effect of freezing on spontaneous, auto-antigen, allergen, and mitogen induced cytokine secretion from peripheral blood mononuclear cells from several groups of patients expressing different cytokine profiles; multiple sclerosis, atopic children, non-atopic children, and pregnant women. The expression of IFN-γ, IL-4, IL-5, IL-9, IL-10, and IL-13 was analysed with ELISA, ELISPOT and/or real time RT-PCR. Our data provide evidence that the process of cryopreservation and thawing does affect the expression of cytokines, both at the protein and the mRNA level. Moreover, the effect varied among different cytokines, different stimuli, and different patient groups, which partly may be explained by differences in optimal freezing conditions for non-activated and activated cells. An increase of allergen and PHA stimulated IFN-γ secretion in atopic children was found following cryopreservation, but no such increase in auto-antigen induced IFN-γ was seen in MS-patients. The most consistent finding was that expression of IL-4 was generally decreased in spontaneous and auto-antigen/allergen induced expression in cryopreserved cells. In conclusion, this study points out the importance of investigation of the effects of freezing for each cytokine, stimuli and patient group before using frozen cells in studies of in vitro cytokine secretion.
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| 2. |
- Dzidic, Majda, et al.
(författare)
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Allergy development is associated with consumption of breastmilk with a reduced microbial richness in the first month of life
- 2020
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Ingår i: Pediatric Allergy and Immunology. - : WILEY. - 0905-6157 .- 1399-3038. ; 31, s. 250-257
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Tidskriftsartikel (refereegranskat)abstract
- Background Early colonization with a diverse microbiota seems to play a crucial role for appropriate immune maturation during childhood. Breastmilk microbiota is one important source of microbes for the infant, transferred together with maternal IgA antibodies. We previously observed that allergy development during childhood was associated with aberrant IgA responses to the gut microbiota already at 1 month of age, when the IgA antibodies are predominantly maternally derived in breastfed infants. Objective To determine the microbial composition and IgA-coated bacteria in breastmilk in relation to allergy development in children participating in an intervention trial with pre- and post-natal Lactobacillus reuteri supplementation. Methods A combination of flow cytometric cell sorting and 16S rRNA gene sequencing was used to characterize the bacterial recognition patterns by IgA in breastmilk samples collected one month post-partum from 40 mothers whose children did or did not develop allergic and asthmatic symptoms during the first 7 years of age. Results The milk fed to children developing allergic manifestations had significantly lower bacterial richness, when compared to the milk given to children that remained healthy. Probiotic treatment influenced the breastmilk microbiota composition. However, the proportions of IgA-coated bacteria, the total bacterial load and the patterns of IgA-coating were similar in breastmilk between mothers of healthy children and those developing allergies. Conclusion Consumption of breastmilk with a reduced microbial richness in the first month of life may play an important role in allergy development during childhood.
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| 3. |
- Permert, Johan, et al.
(författare)
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Life Science på östgötska : förslag till Life science-satsning i Östergötland.
- 2018
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Rapport (övrigt vetenskapligt/konstnärligt)abstract
- Östergötland är inte någon traditionell Life Science-nod i Sverige, men beslutade våren 2017 att undersöka möjligheterna att finna en position baserat på erkänd kunskapsbas kring mötet människa – teknik. En förstudie initierades med Region Direktören som beställare och uppdraget innebar att kartlägga, analysera och ge förslag till hur en Life Science-satsning skulle kunna vara genomförbar i Östergötland. Triple Helix-modellen, det vill säga samhandling mellan offentlig sektor, universitet och näringsliv är den anmodade modellen för Life Science-satsningar i Sverige. Den tidigare Life Science-satsningen i Östergötland hade fallit på grund av obalans i styrka hos Triple Helixens ingående parter.Ett lyckat Triple Helix-initiativ i Östergötland, som även fått internationell genomslagskraft är CMIV, där radiologin är världskänt genom samarbetet mellan RÖ, LiU och Sectra. Detta initiativ är ett exempel på Triple Helix-samhandling som gett resultat för alla ingående parter. Runt CMIV kan såväl samhällsnytta som patientnytta och tillväxt tydligt identifieras. Ytterligare ett exempel som lyftes fram i direktivet är utveckling av kliniska beslutsstöd för användning i klinisk verksamhet och vidare forskning på kliniska data.För att få en rik bild av CMIV, och andra initiativ kring Life Science i Östergötland, har arbetsgruppen använt sig av berättarteknik och på djupet studerat fem fall inom Life Science-området. Analys av dessa fall har sedan jämförts med erfarenheter av Life Science från andra regioner och ett förslag till lösning har successivt växt fram i diskussioner med arbetsgruppen och den taktiska styrgruppen. Triple Helix-samhandling är tämligen utmanande i praktiken då de olika aktörerna i vård, forskning- och näringsliv måste samhandla för att uppnå ett gemensamt mål. Berättelserna vittnar om att det krävs vilja, mod och förmåga att korsa såväl ämnes- som organisatoriska gränser. Slutsatsen är därför att det finns behov av att träna denna förmåga, men också att skapa en ”en väg in” där möten kan uppstå för att identifiera, matcha och förfina initiativ som kan lösas med hjälp av Triple Helix-samhandling.Förstudien visar att det finns goda förutsättningar att genomföra en Life Science-satsning i Östergötland. Ett flerfakultetsuniversitet samt ett komplett sjukvårdssystem ger den mylla av kunskap, problem/behov samt kritiska massa som visat sig krävas för den här typen av satsningar. Att begränsa den möjligheten till enbart vidareutveckling av CMIV och byggandet av kliniska beslutsstöd vore dock inte att göra möjligheterna rättvisa. Istället föreslås två interrelaterade verksamheter vars huvuduppdrag är att facilitera och stödja innovation och utveckling inom Life Science-sektorn i Östergötland; Triple Helix-Labb och Triple Helix-Akademi. I Triple Helix-Labbet kan problem/behov och lösningar mötas, matchas och förfinas i en vägledningsprocess som kan leda till såväl ökad patientnytta som ökad tillväxt i Östergötland. I Triple Helix-Akademin stärks förmågan till samhandling i Triple Helix för att överbyggakunskap, förståelse och respekt för de värdesystem som korsas. Genom dessa interorganisatoriska strukturer finns förutsättningar att adressera vårdens problem, samtidigt som dessa kan agera tillväxtmotor i Östergötland. I förstudien identifieras fyra olika växtvägar som kan stimuleras via ovan nämna strukturer.För att få utväxling av ovanstående förslag behöver strukturer med närliggande och överlappande uppdrag ses över och ersättas/integreras i Triple Helix-Labbet. Detta arbete är påbörjat i förstudien, men behöver förfinas i det etableringsuppdrag som är nästa steg för att kraftsamla kring Life Science i Östergötland. En Life Science-satsning är en långsiktig handling och kräver en politisk överenskommelse för att bli hållbar över tid. Därför ses detta som en förutsättning för att starta det etableringsprojekt som föreslås i föreliggande rapport.Den här förstudien handlar om Östergötland. Redan idag finns dock etablerade samarbeten med Sydöstra sjukvårdsregionen men även nationellt och internationellt. Östergötland är därmed en nod i flera större nav, beroende på vilket perspektiv som antas. Nodtänkandet är en av förstudiens viktigaste grundpelare, men samtidigt måste förändringsarbetet starta hos var och en av de ingående parterna i en Life Science-satsning och därmed adresserar förstudien främst samhandling mellan de parter som utgör själva hjärtat i satsningen; RÖ och LiU.Den största risken för att en Life Science-satsning ska fallera även denna gång är att den politiska överenskommelsen uteblir, eller att RÖ och LiU stannar i sina invanda samverkanspositioner, och därmed inte antar samhällsutmaningen om ökad patientnytta och ökad tillväxt.Förstudien beslutades av Regionstyrelsen 2018-11-08.
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| 4. |
- Svenvik, Maria, 1973-, et al.
(författare)
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Plasma oxylipin levels associated with preterm birth in preterm labor✰
- 2021
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Ingår i: Prostaglandins, Leukotrienes and Essential Fatty Acids. - : Elsevier. - 0952-3278 .- 1532-2823. ; 166
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Tidskriftsartikel (refereegranskat)abstract
- Introduction Preterm labor is a common clinical problem in obstetrics. Since the majority of women with preterm labor eventually deliver at full term, biomarkers are needed to more accurately predict who will deliver preterm. Oxylipins, given their importance in inflammation regulation, are highly interesting in this respect since labor is an inflammatory process.Methods Eighty women with preterm labor before 34 weeks of gestation were enrolled in a prospective observational multi-center cohort study. Oxylipin levels of 67 analytes in plasma samples were analyzed by liquid chromatography coupled to tandem mass spectrometry.Results Twenty-one (26%) of the women delivered before 34 weeks of gestation, and of those women, fourteen delivered within 48 h of admission. Logistic multivariate regression showed that lower levels of 9,10-DiHODE were associated with delivery before 34 weeks of gestation (aOR 0.12 (0.024–0.62)) and within 48 h ((aOR 0.13 (0.019–0.93)). Furthermore, higher levels of 11,12-DiHETrE were associated with delivery before 34 weeks of gestation ((aOR 6.19 (1.17–32.7)) and higher levels of 8-HETE were associated with delivery within 48 h ((aOR 5.01 (1.13–22.14)).Conclusions The oxylipin 9,10-DiHODE may be protective in preterm labor, both for delivery after 34 weeks of gestation and for delivery later than 48 h of admission, whereas 11,12-DiHETrE and 8-HETE display the opposite effect. Larger studies are needed to validate these mediators as biomarkers for prediction of preterm birth following preterm labor.
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| 5. |
- Svenvik, Maria, 1973-
(författare)
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Prediction of Spontaneous Preterm Birth : Clinical and Immunological Aspects
- 2021
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Background: Preterm birth (PTB), defined as birth before 37 weeks of gestation, accounts for most neonatal death and morbidity. Accurate prediction is a pre-requisite for the prevention and proper management of PTB. However, methods for prediction are unsatisfactory, although sonographic cervical length has a moderate predictive value. For clinical utility, adding a biomarker could increase the predictive accuracy. The immune system has an important regulatory role during pregnancy. Thus, presumptive predictive biomarkers may be searched for among immune-related molecules, such as cytokines, chemokines and other inflammation-associated mediators. The aims of this thesis were to identify clinical risk factors and immunological prediction markers for PTB, both in women at increased risk of PTB because of preterm labour (PTL) or preterm prelabour rupture of the membranes (PPROM), and in asymptomatic women in early pregnancy. An additional aim was to explore immune reaction patterns in PTL and PPROM compared to normal pregnancy. Material and methods: In a retrospective registry study, including 20,643 women who delivered during a five-year period, risk factors for Apgar score <7 at five minutes and risk factors for PTB <32 weeks were investigated using univariate and multivariate logistic regression. Furthermore, in a multi-centre mixed case-control and prospective cohort study of women with PTL <34 weeks of gestation (n=80), PPROM (n=40), as well as antenatal controls (n=44) and controls in labour at term (n=40), plasma levels of cytokines and chemokines representing different types of immune responses were analysed with a multiplex bead assay. In addition, an extended protein analysis exploring 92 inflammation-associated plasma proteins using proximity extension assay (PEA) was performed, as well as analysis of 67 different oxylipins by liquid chromatography coupled to tandem mass spectrometry. Finally, the PEA technique was used also to explore plasma proteins in a case-control study including 46 women with PTB and 46 women with normal pregnancies and delivery at term. Results and conclusions: A number of partly preventable clinical risk factors for PTB <32 weeks were identified, for example smoking (odds ratio (OR) 1.61 (95% confidence interval (CI) 1.07-2.41)); preeclampsia (OR 5.48 (95% CI 3.39-8.86)); and multiple gestation (OR 15 (95% CI 10-24)). The most evident risk factor for low Apgar scores was PTB; the more preterm the higher the risk. This provides important information for health care professionals, and offers motivations for preventive strategies regarding smoking cessation. Both PTL and PPROM were associated with a more pro-inflammatory profile compared to antenatal controls, with an increase in CXCL1. In addition, PTL showed higher CCL17 levels, and PPROM showed higher IL-6 levels compared with normal pregnancy. The inflammatory profile was even higher in labour at term, reflected by higher levels of CXCL1, CXCL8, and IL-6 compared with PTL, probably due to the more advanced stage of the parturition process in these women. To identify women with PTL and subsequent PTB <34 weeks of gestation, we found that a combination of the proteins IL-6, IL-17C, IL-10RB, and FGF-23 strongly correlated with PTB <34 weeks with an area under the curve (AUC) of 0.90; inferring a sensitivity of 90%, and specificity of 74%. For the prediction of delivery within 48 hours in women with PTL, the combination of IL-6 and IL-17C displayed an AUC of 0.88, with a sensitivity of 100%, and a specificity of 71%. Additionally, plasma levels of oxylipins were associated with time of birth. Lower levels of 9,10-DiHODE were associated with PTB <34 weeks (adjusted (a) OR 0.12 (0.024-0.62)) and with delivery within 48 hours (aOR 0.13 (0.019-0.93)), while higher levels of 11,12-DiHETre were associated with delivery <34 weeks (aOR 6.19 (1.17-32.7)) and higher levels of 8-HETE were associated with delivery within 48 hours (aOR 5.01 (1.13-22.14)). In asymptomatic women, plasma levels of combinations of inflammation-associated proteins in the first and the second trimester also revealed predictive information regarding subsequent risk for PTB <34 weeks. Combining MMP10trim1, sCD40trim2, MCSFtrim2, Flt3Ltrim2, and FGF-21diff (diff= difference in protein levels comparing the first and second trimesters) provided a prediction model with an AUC of 0.90. Proteins from the first trimester exclusively (sCD40 and MMP10) rendered an AUC of 0.76. This work provides valuable knowledge in the field of PTB and PTL with useful information on risk factors for PTB. Important associations between levels of inflammation-associated proteins and oxylipins with PTB following PTL were found. Before these findings can have clinical implications, they need to be validated in other cohorts. Additionally, in order to be clinically useful as a prediction tool for PTB, a bedside test is needed. Since the PEA technique is PCR-based, this might be achievable. For prediction of PTB in early pregnancy, we have interesting findings with acceptable accuracy based on samples from both the first and the second trimesters. However, as preventive interventions for PTB are preferably initiated early in pregnancy, a prediction tool has better value if it is based on plasma samples from the first trimester. Therefore, we plan to extend this study and evaluate other potential protein biomarkers.
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| 6. |
- Ahlbeck, Lars, 1964-, et al.
(författare)
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Intralymphatic allergen immunotherapy against pollen allergy. A 3-year open follow-up study of 10 patients
- 2018
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Ingår i: Annals of Allergy, Asthma & Immunology. - : Elsevier. - 1081-1206 .- 1534-4436. ; 121:5, s. 626-627
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Tidskriftsartikel (refereegranskat)abstract
- To date, allergen immunotherapy (AIT) is the only treatment that affects the long-term development of allergic rhinoconjunctivitis and induces clinical tolerance primarily by stimulating regulatory T (Treg) cells, attenuating T helper 2 (Th2) responses and synthesis of blocking antibodies1. Conventional AIT with subcutaneous injections, sublingual tablets or drops is effective, but consumes time and resources 2.
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| 7. |
- Ahlbeck, Lars, 1964-
(författare)
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Intralymphatic Immunotherapy : A Novel Route to Ameliorate Allergic Rhinitis Due to Pollen
- 2024
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Allergy to pollen and animal dander is a major public health problem. Close to 30% of the population have symptoms from the upper and/or lower respiratory tract when they meet fur animals or pollen. Whereas symptom-relieving medications have a good to sufficient effect on about 80% of those affected, a large group of 10–20% have severe symptoms, despite medication, with an impact on well-being and ability to work. In Sweden, the annual cost of allergy was calculated at €1.3 billion in 2014.Immunotherapy is effective in treating and preventing pollen allergy and allergic asthma, but is expensive, complicated, requiring 40 injections, and takes more than three years to complete if subcutaneous injections are used. Tablets placed under the tongue are another method, with one tablet taken every day for three years. Only 1.5‰ receive such treatment, yet just over 3% would need it.With intralymphatic immunotherapy, a small dose of allergen is given in a lymph node in the groin on 3 occasions, one month apart. As this method takes only eight weeks, it is a much faster and less costly treatment. However, although several studies have shown that the treatment is safe, its efficacy remains the subject of doubt.Our pilot study in 2012, with a 3-year follow-up to 2015, showed encouraging results, and was followed by a double-blind randomised study with 72 participants from 2014 to 2018. The research subjects then received treatment with birch and grass pollen extract or one extract and a placebo. Regardless of treatment, symptoms, quality of life and medication consumption improved during the birch and grass pollen seasons in the 3 years after treatment. Increased frequencies of T-regulatory lymphocytes may explain the non-specific effects.In 2017 to 2018, we conducted a double-blind study with 38 participants, half of whom received placebo and half, active treatment. In this study, we saw no difference between the treatment groups in the first year after treatment. However, after discontinuation and unblinding in 2019, i.e., two years after treatment, the actively treated group improved in terms of symptoms, and quality of life was improved compared with the placebo group despite less need for medication. T-regulatory lymphocytes increased one year after treatment only in the actively treated group.A long-term follow-up of the research subjects from our two larger studies in 2022, i.e., five to eight years after treatment, showed in the double-blind study without a pure placebo that the scores for symptoms, medication use, and quality of life remained as low as after the first three years. In the placebo-controlled study, a statistically significant improvement in symptoms remained during the grass pollen season. Analysing the two studies together, symptom improvement was significant even during the birch pollen season. Thus, although the effect does not seem to diminish, those who did not receive birch, but only grass, needed to use more medication during the birch pollen season in 2022, seven to eight years after treatment. Moreover, those who did not receive grass but only birch needed more medication during the grass pollen season. This may suggest that the non-specific effect begins to wane after seven to eight years.Allergy to pollen is a major problem for individuals and society, where symptom-relieving treatment with drugs is not enough for many. They can be helped with immunotherapy, which takes at least three years, is expensive and fraught with side effects. In contrast, intralymphatic immunotherapy involves three injections over eight weeks. Our three studies show that the treatment is safe and indicate that it has a clinical effect up to eight years after treatment. T-regulatory cells appear to be important to the immunological mechanism, leading to tolerance to pollen.
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| 8. |
- Ahlbeck, Lars, 1964-, et al.
(författare)
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Intralymphatic immunotherapy with one or two allergens renders similar clinical response in patients with allergic rhinitis due to birch and grass pollen
- 2022
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Ingår i: Clinical and Experimental Allergy. - Chichester, United Kingdom : Wiley-Blackwell Publishing Inc.. - 0954-7894 .- 1365-2222. ; 52:6, s. 747-759
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Tidskriftsartikel (refereegranskat)abstract
- IntroductionThere is a need for a fast, efficient and safe way to induce tolerance in patients with severe allergic rhinitis. Intralymphatic immune therapy has been shown to be effective. MethodsPatients with severe birch and timothy allergy were randomized and received three doses of 0.1 ml of birch and 5-grass allergen extracts (10,000 SQ units/ml, ALK-Abello), or birch and placebo or 5-grass and placebo by ultrasound-guided injections into inguinal lymph nodes at monthly intervals. Rhinoconjunctivitis total symptom score, medication score and rhinoconjunctivitis quality of life questionnaire were evaluated before treatment and after each birch and grass pollen season during three subsequent years. Circulating proportions of T helper subsets and allergen-induced cytokine and chemokine production were analysed by flow cytometry and Luminex.Results The three groups reported fewer symptoms, lower use of medication and improved quality of life during the birch and grass pollen seasons each year after treatment at an almost similar rate independently of treatment with one or two allergens. Mild local pain was the most common adverse event. IgE levels to birch decreased, whereas birch-induced IL-10 secretion increased in all three groups. IgG4 levels to birch and timothy and skin prick test reactivity remained mainly unchanged. Conjunctival challenge tests with timothy extract showed a higher threshold for allergen. In all three groups, regulatory T cell frequencies were increased 3 years after treatment.Conclusions Intralymphatic immunotherapy with one or two allergens in patients with grass and birch pollen allergy was safe, effective and may be associated with bystander immune modulatory responses.
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| 9. |
- Bensberg, Maike, 1993-
(författare)
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DNA methylation in T cell leukaemia
- 2024
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- T cell acute lymphoblastic leukaemia (T-ALL) is a predominantly paediatric cancer that stems from malignant transformation of developing T cells. While the disease has an overall survival rate of 80%, the intense chemotherapy treatment causes severe toxicity and long-term side effects. Furthermore, the survival rate for patients in relapse is less than 25%. Consequently, there is a need for improved therapy options to reduce treatment-related side effects and improve the survival rate of relapsed patients. Targeting aberrant DNA methylation with hypomethylating agents (HMAs) has been successful in the treatment of myelodysplastic syndromes and acute myeloid leukaemia but has not been routinely used in the treatment of T-ALL, despite DNA hypomethylation being observed in T-ALL patients. In this work, we employed a comprehensive set of molecular and sequencing-based techniques to explore the possibilities of HMAs as a treatment option for T-ALL.We made the discovery that the DNA demethylating enzyme ten-eleven translocation 2, TET2, is downregulated or completely silenced in primary T-ALL. Moreover, the TET2 promoter was highly methylated in a group of patients, suggesting that TET2 itself can be silenced through DNA methylation in T-ALL. By treatment with HMAs, TET2 was demethylated in T-ALL cell lines and was one of few genes that was activated upon loss of DNA methylation, indicating that TET2 expression is regulated by DNA methylation in T-ALL cell lines. The development of a novel HMA, the DNMT1-specific inhibitor GSK-3685032, offers a tool to reveal the mechanism of action of the traditional HMAs, 5- azacytidine and decitabine, and to study the effects of acute loss of DNA methylation on cancer cells. We found that 5-azacytidine and decitabine are cytotoxic to T-ALL cells primarily by creating DNA double strand breaks. In contrast, GSK did not prompt a DNA damage response and instead reduced global DNA methylation to as little as 18% with limited cytotoxicity only occurring after levels of DNA methylation had dropped below 30%, a level of demethylation not achieved with DEC or AZA.T-ALL is more than two times more common in boys than girls and mutations in X-linked tumour suppressor genes that escape X inactivation, have been suggested as an underlying cause for the observed sex-bias. In theory, these aberrations would be more detrimental in XYmale cells than in XX-female cells due to the presence of an extra protective copy of the gene in females. We profiled DNA methylation during T cell development and created a map of sex-specific gene expression and expression from the inactive X chromosome, finding that some, but not all, suggested tumour suppressor genes in fact escape X inactivation. These results highlight the importance of profiling the healthy cells that T-ALL arises from to correctly judge the functional impact of gene dysregulation in cancer.In the last study, we aimed to investigate the role of N6-adenine methylation (6mdA) during T cell differentiation. While 6mdA is common in bacteria it is much rarer in humans. Nevertheless, 6mdA has previously been associated with several cellular processes, including cancer progression. Our study calls the presence of 6mdA in mammals into question by exposing limitations of the techniques used in its analysis. We show that contamination with bacterial DNA or 6mAcontaining RNA, nonspecific antibody binding, and low precision of third-generation sequencing techniques all hinder the detection and investigation of rare DNA modifications, such as 6mdA.Together, this work is an in-depth study of the function and the potential of DNA methylation in the biology of healthy and malignant T cells.
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| 10. |
- Bruno, Valentina, 1986-
(författare)
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Clinical and immunological aspects on recurrent pregnancy loss
- 2020
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Paper I. Effects of low molecular weight heparin on the polarization and cytokine profile of macrophages and T helper cells in vitro. Sci Rep 2018. In paper I low molecular weight heparin (LMWH) in vitro effects on activation and polarization of central regulatory immune cells, such as Th cells and macrophages, were assessed, since LMWH has been widely used as an empiric treatment in recurrent pregnancy loss (RPL) and its immunological effects are not fully known. Isolated blood monocytes and T helper (Th) cells under different activation and polarizing conditions were cultured without or with LMWH at different concentrations. LMWH exposure induced an activated phenotype of macrophages, with high expression of HLA-DR and CD206 assessed by flow cytometry, associated with increased secretion of Th17-associated CCL20, and decreased secretion of CCL2 (M2-associated) and CCL22 (Th2), as measured by multiplex bead array. In accordance, LMWH exposure to Th cells reduced the proportion of CD25highFoxp3+ regulatory Tcells, and intensified IFN-γ secretion. Collectively, a mainly pro-inflammatory effect was noted on two essential tolerance-promoting cells, suggesting that potential immunological effects of LMWH may be effective mainly at an earlier gestational age to provide an appropriate implantation process in women with recurrent miscarriage. Paper II. Low-molecular-weight-heparin increases Th1- and Th17-associated chemokine levels during pregnancy in women with unexplained recurrent pregnancy loss: a randomized controlled trial. Sci Rep 2019.In paper II we investigated whether LMWH could modulate immune responses in vivo during pregnancy of women with unexplained RPL. A Swedish open multi-centre randomized controlled trial included 45 women treated with tinzaparin and 42 untreated women. Longitudinally collected plasma samples were obtained at gestational weeks (gw) 6, 18, 28 and 34 and analyzed by multiplex bead technology for levels of 11 cytokines and chemokines, chosen to represent inflammation and T-helper subset-associated immunity. LMWH-treated and untreated women showed differences during pregnancy of the Th1-associated chemokines CXCL10 (p = 0.01), CXCL11 (p < 0.001) and the Th17- associated chemokine CCL20 (p = 0.04), while CCL2, CCL17, CCL22, CXCL1, CXCL8, CXCL12, CXCL13 and IL-6 did not differ. Significantly higher plasma levels of CXCL10 and CXCL11 in treated women were detected at gw 28 and 34, compared to the untreated ones. Thus, a potential proinflammatory effect, linked mainly to Th1 immunity, was shown, suggesting an unfavorable effect of LMWH treatment, since Th1 responsea are responsible for breaking the fetal-maternal immune tolerance. Paper III. First-trimester trophoblasts obtained by chorionic villus sampling maintain tolerogenic and proteomic features in successful pregnancies despite a history of unexplained recurrent pregnancy loss. Am J Reprod Immunol. 2020.In paper III we investigate the “local” immune changes in women with RPL, since they potentially could reveal important mechanisms in RPL. Supernatants from superfluous chorionic villus sampling material culture was used in an ex vivo model, to determinate the immune proteomics profile and to perform functional assays for M2 like macrophages and regulatory T cells polarization, assessed by flow cytometry technique. Chorionic villi, human fetally derived placental tissue, were shown to induce an M2 like-phenotype and an expansion of Treg cells in an ex vivo model, and these immunological properties were maintained despite a history of RPL. Accordingly, no differences in the inflammation proteomic profile were found in RPL, compared to controls. Trophoblasts in an ex vivo model thus maintain tolerogenic and proteomic profile features in successful pregnancies, despite a history of RPL.
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