Lymphocyte Immunotherapy Research Articles

Maternal-fetal immunity and recurrent spontaneous abortion.

Recurrent Spontaneous Abortion (RSA) is a common pregnancy complication, that has multifactorial causes, and currently, 40%-50% of cases remain unexplained, referred to as Unexplained RSA (URSA). Due to the elusive etiology and mechanisms, clinical management is exceedingly challenging. In recent years, with the progress in reproductive immunology, a growing body of evidence suggests a relationship between URSA and maternal-fetal immunology, offering hope for the development of tailored treatment strategies. This article provides an immunological perspective on the pathogenesis, diagnosis, and treatment of RSA. On one hand, it comprehensively reviews the immunological mechanisms underlying RSA, including abnormalities in maternal-fetal interface immune tolerance, maternal-fetal interface immune cell function, gut microbiota-mediated immune dysregulation, and vaginal microbiota-mediated immune anomalies. On the other hand, it presents the diagnosis and existing treatment modalities for RSA. This article offers a clear knowledge framework for understanding RSA from an immunological standpoint. In conclusion, while the "layers of the veil" regarding immunological factors in RSA are gradually being unveiled, our current research may only scratch the surface. In terms of immunological etiology, effective diagnostic tools for RSA are currently lacking, and the efficacy and safety of immunotherapies, primarily based on lymphocyte immunotherapy and intravenous immunoglobulin, remain contentious.

Observation on the Efficacy of Lymphocyte Immunotherapy for Recurrent Biochemical Pregnancy Loss

Objective: To implement a lymphocyte immunotherapy program for women experiencing recurrent biochemical pregnancy loss and explore its clinical effects. Methods: The study employed a retrospective design. From January 2015 to January 2016, a total of 100 patients with recurrent biochemical pregnancy loss were enrolled as observation objects. Subsequently, the patients were divided into two groups based on different treatment plans, each comprising 50 cases. The observation group received conventional tocolysis in addition to lymphocyte immunotherapy, while the control group received only conventional tocolysis treatment. Evaluation criteria included pregnancy outcomes, improvements in serum factor levels, and treatment safety, enabling a comparison of the clinical effects of different treatment options. Results: In the observation group, the pregnancy success rate was 82.00%, whereas in the control group, it was 48.00% (χ2 = 12.7033, P < 0.05). Following treatment, levels of interferon-γ (IFN-γ) were higher in the observation group than in the control group, while levels of interleukin-8 (IL-8) and regulatory protein (BANTES) were lower in the observation group compared to the control group (P < 0.05). The difference in treatment safety between the two groups was negligible (P > 0.05). Conclusion: The incorporation of lymphocyte immunotherapy for patients with recurrent biochemical pregnancy loss can enhance successful pregnancy rates, demonstrating significant efficacy and warranting further application.

Systemic immune index predicts tumor-infiltrating lymphocyte intensity and immunotherapy response in small cell lung cancer.

Despite recent progresses in immune checkpoint blockade (ICB) in small-cell lung cancer (SCLC), a lack of understanding regarding the systemic tumor immune environment (STIE) and local tumor immune microenvironment (TIME) makes it difficult to accurately predict clinical outcomes and identify potential beneficiaries from ICB therapy. We enrolled 191 patients with stage I-III SCLC and comprehensively evaluated the prognostic role of STIE by several quantitative measurements, and further integrate it with a local immune score system (LISS) established by eXtreme Gradient Boosting (XGBoost) machine learning algorithm. We also test the value of STIE in beneficiary selection in our independent advanced SCLC cohort receiving programmed cell death 1 ligand 1 (PD-L1) blockade therapy. Among several systemic immune markers, the STIE as assessed by prognostic nutritional index (PNI) was correlated with disease-free survival (DFS) and overall survival (OS), and remained as an independent prognostic factor for SCLC patients [hazard ratio (HR): 0.473, 95% confidence interval (CI): 0.241-0.929, P=0.030]. Higher PNI score was closely associated with inflamed SCLC molecular subtype and local tumor-infiltrating lymphocytes (TILs). We further constructed a LISS which combined top three important local immune biomarkers (CD8+ T-cell count, PD-L1 expression on CD8+ T-cell and CD4+ T-cell count) and integrated it with the PNI score. The final integrated immune risk system was an independent prognostic factor and achieved better predictive performance than Tumor Node Metastasis (TNM) stages and single immune biomarker. Furthermore, PNI-high extensive-stage SCLC patients achieved better clinical response and longer progression-free survival (PFS) (11.8 vs. 5.9 months, P=0.012) from PD-L1 blockade therapy. This study provides a method to investigate the prognostic value of overall immune status by combining the PNI with local immune biomarkers in SCLC. The promising clinical application of PNI in efficacy prediction and beneficiary selection for SCLC immunotherapy is also highlighted.

A comprehensive Bayesian analysis assessing the effectiveness of lymphocyte immunotherapy for recurrent spontaneous abortion

AbstractRecurrent spontaneous abortion (RSA) affects 2%–5% of couples worldwide and remains a subject of debate regarding the effectiveness of lymphocyte immunotherapy (LIT) due to limited retrospective studies. We conducted a comprehensive Bayesian analysis to assess the impact of LIT on RSA. Using data from the Shenzhen Maternity and Child Healthcare Hospital (2001–2020, n = 2316), a Bayesian generalized linear model with predictive projection feature selection was employed. Our analysis revealed a significant improvement in live birth rates for RSA patients undergoing LIT. Notably, LIT had a greater impact compared to the other 85 factors considered. To mitigate research bias, we conducted a Bayesian meta-analysis combining our dataset with 19 previously reported studies (1985–2021, n = 4246). Additionally, we developed an empirical model highlighting the four key factors, which are the LIT result, age, paternal blood type, and anticardiolipin antibody. Younger age (19–27), paternal blood type B, and a positive anticardiolipin antibody (IgM) were associated with better therapeutic outcomes in LIT for RSA. These findings aid clinicians in identifying suitable candidates for LIT and improving treatment outcomes.

The Role of Thoracic Vertebral Body Dosimetry in Minimizing Acute Hematologic Toxicities of Patients With Non-Small Cell Lung Cancer Receiving Lung Radiation Therapy and Immunotherapy

IntroductionHematologic toxicities (HTs) are among the most common toxicities of combined immunotherapy and radiotherapy (RT). It remains essential to prevent RT-induced HTs because they can cause treatment discontinuation (influencing antitumoral effects), and because lymphopenia might dampen the effects of immunotherapy. To date, there are no studies examining the impact of thoracic vertebral body (TVB) RT dose on HTs in non-small cell lung cancer (NSCLC) patients receiving combined lung RT and PD(L)-1 immunotherapy. MethodsFor standardization, all doses were reported as 2-Gy equivalents (EQD2). Mirroring publications prior to the immunotherapy era, TVB volumes referred to T1-T10 and specific dosimetric parameters (DmeanEQD2, V5EQD2‒V60EQD2) were analyzed. Logistic regression estimated associations between grade ≥3 HTs (HT3+) and dosimetric/clinical parameters. Normal tissue complication probability (NTCP) models were constructed by logistic regression analysis modeling for HT3+. Receiver operating characteristic (ROC) analysis delineated TVB dosimetric thresholds, stratification of which was able to evaluate post-RT absolute lymphocyte count (ALC) and immunotherapy responses. Areas under the curve (AUCs) for NTCP models were corroborated by bootstrapping (optimism-corrected) methodology. ResultsIn 132 patients, there were 26 (19.7%) instances of HT3+. On multivariate analysis, DmeanEQD2 and V5EQD2‒V20EQD2 associated with HT3+ (P<0.05 for all). The NTCP models illustrated a 50% probability of HT3+ at a DmeanEQD2=39.8 Gy, V5EQD2=87.4%, V10EQD2=77.0%, and V20EQD2=68.4%. ROC analysis delineated optimal thresholds of HT3+ with DmeanEQD2 ±30.2 Gy, V5EQD2 ±69.1%, V10EQD2 ±64.6%, and V20EQD2 ±53.5%. Patients treated with values above those cutoffs had over double the risk of HT3+, with significant differences in post-RT ALC and immunotherapy responses (P<0.05 for all). AUCs for each individual parameter ranged from 0.743 to 0.798, and combining all four aforementioned cutoffs into a ROC curve resulted in a qualitatively higher AUC (0.836). ConclusionsThis hypothesis-generating work suggests that TVB dosimetry may equate with HT3+ in NSCLC patients undergoing combined lung RT/immunotherapy. Applying TVB dose constraints in this population could reduce HT3+ and avoid dampening of immunotherapy responses, but prospective validation is required.

The effect of lymphocyte immunotherapy (LIT) in modulating immune responses in patients with recurrent pregnancy loss (RPL).

In order to prevent miscarriage in RPL patients, the goal of this study was to determine how well lymphocyte immunotherapy (LIT) works in modifying immunological responses produced by cells, cytokines, transcription factors, and microRNAs. 200 RPL patients and 200 healthy controls were included in the study. Using flow cytometry, it was possible to compare the frequency of cells before and after lymphocyte treatment. Real-time PCR was used to assess the gene expression levels of transcription factors, cytokines, and microRNAs. ELISA method was used to evaluate the level of secretion of cytokines in the serum. Primary evaluation of the immune profile between healthy controls and RPL cases showed a higher frequency of Th17, NK, B cells and a lower frequency of Treg cells in RPL cases. Also, pro-inflammatory cytokines showed increased expression at mRNA and protein levels in the RPL group in comparison with the control group. Whereas, anti-inflammatory cytokines showed decreased expression in RPL patients. Decreased and increased frequency of Th17 and Treg lymphocytes observed in RPL cases following LIT, respectively. The same results obtained for RORγt and FoxP3 mRNA expression as transcription factor of Th17 and Treg cells, respectively. NK cell cytotoxicity decreased after LIT in RPL patients. miR-326a and miR-155 expression after LIT reduced, but miR-146a and miR-10a expression increased in RPL instances. LIT in RPL cases causes to elevation and modulation of anti-inflammatory and pro-inflammatory cytokines. Our data showed that lymphocyte therapy can be proposed as an effective therapeutic agent in RPL patients with immunological background by a modulating inflammatory condition.

Survivin Expression in Luminal Breast Cancer and Adjacent Normal Tissue for Immuno-Oncology Applications.

Survivin (BIRC5) is a tumor-associated antigen (TAA) overexpressed in various tumors but present at low to undetectable levels in normal tissue. Survivin is known to have a high expression in breast cancer (e.g., Ductal Carcinoma in situ (DCIS) and triple negative breast cancer). Previous studies have not compared survivin expression levels in DCIS tumor samples to levels in adjacent, normal breast tissue from the same patient. To ensure the effective use of survivin as a target for T cell immunotherapy of breast cancer, it is essential to ascertain the varying levels of survivin expression between DCIS tumor tissue samples and the adjacent normal breast tissue taken from the same patient simultaneously. Next-generation sequencing of RNA (RNA-seq) in normal breast tissue and tumor breast tissue from five women presenting with DCIS for lumpectomy was used to identify sequence variation and expression levels of survivin. The identity of both tumor and adjacent normal tissue samples were corroborated by histopathology. Survivin was overexpressed in human breast tissue tumor samples relative to the corresponding adjacent human normal breast tissue. Wild-type survivin transcripts were the predominant species identified in all tumor tissue sequenced. This study demonstrates upregulated expression of wild type survivin in DCIS tumor tissue versus normal breast tissue taken from the same patient at the same time, and provides evidence that developing selective cytotoxic T lymphocyte (CTL) immunotherapy for DCIS targeting survivin warrants further study.

Diffuse Pigment Release in a Patient Undergoing Tumor-Infiltrating Lymphocyte Immunotherapy for Acral Malignant Melanoma.

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The Effectiveness of Paternal Lymphocyte Immunotherapy for Recurrent Miscarriage in Couples with Human Leukocyte Antigen Sharing: A Novel Approach

Background: The aetiology of recurrent pregnancy loss (RPL) is varied and ranges from genetic abnormalities, autoimmune, uterine structural abnormalities, thrombophilic disorders, endocrinologic dysfunction, infective to idiopathic factors. Reproductive immunology may provide an area of opportunity in treatment of idiopathic cases. Research has indicated that any amount of HLA compatibility among spouses leads to immunological perturbations leading to higher RPL rates. These disturbances in alloimmune parameters are found to be significantly reduced after a successful immunotherapy with paternal lymphocytes immunotherapy (LIT) among couples who share HLA. Aim: To analyze the role of alloimmune factors in couples who are considered unexplained RPL by testing HLA sharing between the partners and to determine the effect of lymphocyte immunotherapy (LIT) on live birth rate in couples with HLA sharing. Methods: This retrospective observational study was conducted in a single tertiary center in Bangalore for a duration of three years. Couples who satisfied the inclusion and exclusion criteria were selected and HLA sharing between the partners was tested. Couples with HLA sharing received LIT before and during pregnancy. The pregnancy and live birth rates were calculated and compared with couples with HLA sharing who did not receive LIT. Results: Out of the 199 couples who were screened for HLA sharing among partners, 146 couples had different degrees of HLA sharing. 81 couples received LIT and 32 did not opt for LIT and were taken as control group. The pregnancy and live birth rates were significantly higher in the LIT group compared to control group (77.7% vs 40.6%, p-0.0001, OR 5.1, 95% CI 2.10-11.4 and 56.7% vs 21.8%, p-0.0002, OR 4.6, 95% CI 2.13-13.8 respectively). Miscarriage rates were similar between the two groups. Conclusion: Partner lymphocyte immunotherapy is a novel treatment option in improving the pregnancy outcomes among women with unexplained RPL and HLA sharing among partners.

Contradictory Effect of Lymphocyte Therapy and Prednisolone Therapy on CD3+CD8+CD56+ Natural Killer T Population in Women with Recurrent Spontaneous Abortion.

Natural killer T (NKT) cells are influential immune cells in pregnancy failures, including recurrent spontaneous abortion (RSA). Different approaches are used for these disorders due to their effects on maternal immunomodulation. In the present study, we compared the effects of two typical immunotherapies (lymphocyte immunotherapy [LIT] and low-dose prednisolone) on CD3+CD56+CD16+ and CD3+CD56+CD8+ cells as two distinct subsets of NKT cells in Women with RSA. This study was a comparative cohort study conducted from 2021 to 2022. One hundred and five women with RSA were distributed into three treatment groups randomly. Fifty women in the group of low-dose prednisolone therapy, fifty women in the LIT group and five women without any treatment as the control group were included in the study. NK and NKT cell subsets were assessed using flow cytometry. Furthermore, the concentration of interferon-gamma (IFN-γ), transforming growth factor-beta (TGF-β) and interleukin-10 (IL-10) was measured quantitatively using the enzyme-linked immunosorbent assay technique. Normality and comparisons between study groups were performed by non-parametric unpaired Mann-Whitney, Kruskal-Wallis rank sum test, and one-way ANOVA. The percentage of CD56dim NK cells was increased after prednisolone therapy, while this population significantly decreased in the LIT group. In contrast to the LIT group, the administration of prednisolone increased CD3+CD8+CD56+ NKT cells (P < 0.0001), which is helpful for pregnancy. The effect of the investigated treatment approaches on the population of peripheral CD3+CD56+CD16+ NKT cells of women with RSA was not adequately significant. The same situation was also observed regarding the serum level of IFN-γ. However, a significant decrease in serum levels of IL-10 and TGF-β was observed after prednisolone therapy. The lower capability of LIT in changing the population of NKT cells compared to prednisolone therapy may be due to its mechanism of action, which is related to the production of blocking antibodies. These treatment approaches had different effects on NKT cells, indicating that NKT cell population and function can be affected using LIT and prednisolone therapy distinctly. In addition, prednisolone therapy and LIT in women with normal serum levels of IFN-γ have no harmful effects in changing the production of this critical cytokine.

Prospective study of adoptive activated αβT lymphocyte immunotherapy for refractory cancers: development and validation of a response scoring system

Background aimsThis prospective clinical study aimed to determine the efficacy and prognostic factors of adoptive activated αβT lymphocyte immunotherapy for various refractory cancers. The primary endpoint was overall survival (OS), and the secondary endpoint was radiological response. MethodsThe authors treated 96 patients. Activated αβT lymphocytes were infused every 2 weeks for a total of six times. Prognostic factors were identified by analyzing clinical and laboratory data obtained before therapy. ResultsMedian survival time (MST) was 150 days (95% confidence interval, 105–191), and approximately 20% of patients achieved disease control (complete response + partial response + stable disease). According to the multivariate Cox proportional hazards model with Akaike information criterion–best subset selection, sex, concurrent therapy, neutrophil/lymphocyte ratio, albumin, lactate dehydrogenase, CD4:CD8 ratio and T helper (Th)1:Th2 ratio were strong prognostic factors. Using parameter estimates of the Cox analysis, the authors developed a response scoring system. The authors then determined the threshold of the response score between responders and non-responders. This threshold was able to significantly differentiate OS of responders from that of non-responders. MST of responders was longer than that of non-responders (317.5 days versus 74 days). The validity of this response scoring system was then confirmed by internal validation. ConclusionsAdoptive activated αβT lymphocyte immunotherapy has clinical efficacy in certain patients. The authors’ scoring system is the first prognostic model reported for this therapy, and it is useful for selecting patients who might obtain a better prognosis through this modality.

Efficacy of lymphocyte immunotherapy in the treatment of recurrent pregnancy loss from alloimmunity: A systematic review and meta-analysis.

The efficacy of lymphocyte immunotherapy (LIT) in the treatment of recurrent pregnancy loss (RPL) from alloimmunity has been debated for years. There is conflicting evidence on the therapeutic role of LIT, since the etiology of most cases of RPL is previously classified as idiopathic. A systematic search of PubMed and Cochrane databases was done for randomized controlled trials that assessed the efficacy of paternal lymphocyte or third donor LIT among patients with primary or secondary RPL. The primary outcome was live birth rate after LIT. Random-effect meta-analysis was conducted using the software RevMan 5.4. Pre-planned subgroup analyses of source of lymphocytes, timing and frequency of administration, and concentration per immunization dose were conducted. Data from eight trials showed a statistically significant benefit of LIT (RR=1.45, 95% CI 1.05-2.01). The overall live birth rate is higher in the treatment group (65.6%) compared to placebo or no treatment (45.2%). Subgroup analysis based on source of lymphocytes revealed a trend towards benefit with paternal LIT but with wide confidence interval (RR=1.34, 95% CI=.84-2.14). Multiple doses of immunotherapy before pregnancy and low dose (5×106 cells) LIT showed significant benefit. Sensitivity analysis involving studies with a low risk of bias demonstrated significant benefit of increased live birth rate among patients treated with LIT compared to those who received placebo or no treatment (RR=1.97, 95% CI=1.53-2.53). LIT demonstrate benefit in improving pregnancy outcome of patients with RPL from alloimmunity.

Adverse drug reactions following lymphocyte immunotherapy for the treatment of infertility: A retrospective study.

Unexplained infertility is a major burden for couples who want to have children. Lymphocyte immunotherapy (LIT) could be a therapeutic help for these couples. Although LIT has been carried out for decades, the data on the success of therapy are still controversial and there is hardly information on possible adverse drug reactions. In this study, we used a questionnaire to determine the frequency of local and systemic adverse drug reactions in our patients who were treated with LIT between 2017 and 2020 (n=302). In addition, we asked about pregnancies and/or live births after LIT in a 2-year follow-up (n=140). Most of the patients reported the occurrence of mild local adverse drug reactions in a period of less than 4 weeks: Over 75% reported moderate erythema, itching or swelling, over 10% erythema, itching or swelling as more pronounced adverse drug reaction. Blistering was specified in 10% of the cases. Serious adverse drug reactions or adverse events were not described. In the follow-up, 69% of our patients stated a pregnancy after LIT, and 50% a life birth. Overall, LIT represents a well-tolerated therapy for couples with unexplained infertility, however, more evidence is needed on the benefits.

WHOLE LUNG LAVAGE IN A CASE OF PULMONARY ALVEOLAR PROTEINOSIS

TOPIC: Procedures TYPE: Global Case Reports INTRODUCTION: Pulmonary Alveolar Proteinosis (PAP) is a syndrome characterized by progressive accumulation of surfactant phospholipids and proteins within alveoli and terminal airways due to decreased surfactant clearance by alveolar macrophage. There are three types,1) congenital2)primary/idiopathic 3) secondary Open lung biopsy with PAS stain is gold standard for diagnosis. Other supportive investigation are chest x ray, HRCT Thorax, GM CSF antibody, hypoxia in ABGA, restrictive or normal PFT.Whole Lung Lavage is mainstay of treatment. GM CSF, plasmapheresis, lung transplantation, supportive care, anti B lymphocyte immunotherapy are other modes of treatment. Some cases shows spontaneous remission. Majority of patients do not require second cycle of WLL. CASE PRESENTATION: 30 year, male, married, non addicted, no co morbidity patient presented to Department of Pulmonary Medicine, B J Medical College, Civil Hospital, Ahmedabad, with chief complaint of progressive difficulty in breathing over 6 to 8 months and dry cough with occasionally whitish expectoration.He works at ship breaking yard.He came with HRCT thorax which had "crazy paving” pattern. On examination he had bilateral fine crepitation and Spo2 86% at room air which reduce to 82% on exertion. So patient hospitalized for further investigations, diagnosis and treatment. DISCUSSION: After admission all routine blood investigations carried out which were within normal limit. On ABGA at room air PaO2 was 66 mmHg. 2D Echo was normal. On PFT restrictive pattern seen. RT PCR for COVID 19 was negative. After exclusion of other causes for HRCT pattern and clinical symptoms bronchoscopy was performed. on bronchoscopy white, frothy proteinosis material seen. On bronchial wash amorphous PAS stained material seen. On culture and sensitivity no organism were cultivated. So after confirmation of diagnosis of pulmonary alveolar proteinosis we planned for whole lung lavage. From HRCT thorax left lung was more involved so on first sitting left sided whole lung lavage performed. With help of left sided double lumen endotracheal tube and single lung ventilation we carried out lavage in 11 sequential cycle. Each cycle took approximately 15-20 minutes. Post procedure patient mechanically ventilated for 16 hours and then extubated without any complication. Post procedure patient improved clinically and radiologically. His right side whole lung lavage is planned after a month. Prior to this whole lung lavage, one patient's WLL was done at our institute. CONCLUSIONS: PAP is relatively rare disease with estimated to be 3.7 case per million individuals. For diagnosis open lung biopsy is the gold standard. Whole lung lavage is mainstay of treatment. This procedure require well pre procedure planning and co ordination among pulmonologist, anesthetist and physiotherapist. REFERENCE #1: Fishman's pulmonary diseases and disorders REFERENCE #2: DeMello De, Lin Z: pulmonary alveolar proteinosis: a review. Pediatr Pathol Mol Med 20:413-432, 2001 REFERENCE #3: Whole lung lavage - technical details, challenges and management of complications. Ahmed Awab, Journal of Thoracic Disease 2017;9(6):1697-1706 DISCLOSURES: no disclosure on file for Ghanshyam Borisagar; no disclosure on file for AMIT DEDUN; No relevant relationships by rajdeep dhandhukiya, source=Web Response No relevant relationships by RAJESH SOLANKI, source=Web Response

Live Birth Rates after Active Immunization with Partner Lymphocytes.

Although many potential causes have been established for recurrent implantation failure (RIF) and recurrent miscarriage (RM), about 50% of these remain idiopathic. Scientific research is focused on immunological risk factors. In the present study, we aim to evaluate live birth rates after immunization with paternal lymphocytes (lymphocyte immunotherapy (LIT)). This retrospective study consisted of 148 couples with a history of RM and/or RIF. The women underwent immunization with lymphocytes of their respective partners from November 2017 to August 2019. Fifty-five patients (43%) had live births. Stratified by indication (RM, RIF, combined), live birth rates in the RM and the combined group were significantly higher than that in the RIF group (53%, 59% and 33%, respectively, p = 0.02). The difference was especially noticeable during the first 90 days after immunization (conception rate leading to live births: 31%, 23% and 8% for RM, the combined group and RIF, respectively; p = 0.005), while there was no difference between groups during the later follow-up. LIT was associated with high live birth rates, especially in women with recurrent miscarriage. In view of the limited data from randomized studies, LIT cannot be recommended as routine therapy. However, it may be considered in individual cases.

Influence of Bushen Tiaochong Cycle Therapy on Th1/Th2 Deviation, Sex Hormone Level, and Pregnancy Outcome of Alloimmune Recurrent Spontaneous Abortion.

Objective To analyze the influence of Bushen Tiaochong cycle therapy on Th1/Th2 deviation, sex hormone level, and pregnancy outcome of alloimmune recurrent spontaneous abortion (RSA). Methods From August 2018 to April 2020, 130 patients with alloimmune RSA who met the inclusion criteria were randomly divided into the control group (n = 65) and the study group (n = 65). The former received lymphocyte immunotherapy (LIT), and the latter received Bushen Tiaochong cycle therapy on the basis of LIT. The treatment ended at 12 w of pregnancy. The total score of traditional Chinese medicine (TCM) syndromes, Th1 cytokine (IL-2), Th2 cytokine (IL-10), and related hormones (chorionic gonadotropin (hCG) and progesterone (P)) were compared between the two groups before and after treatment. The positive rate of blocking antibody (BA), pregnancy success rate, and preterm birth rate were counted. Result After treatment, the total score of TCM syndromes, IL-2 level, and Th1/Th2 ratio in the two groups decreased significantly, while the levels of IL-10, hCG, and P increased significantly, and the study group improved significantly compared with the control group (P < 0.05). The positive rate of BA and pregnancy success rate in the study group were higher than those in the control group (P < 0.05). There was no significant difference in the preterm birth rate between the two groups (P > 0.05). Conclusion On the basis of routine western medicine treatment, a combined application of Bushen Tiaochong cycle therapy can significantly improve the Th1/Th2 deviation, serum sex hormone level, and pregnancy outcome in patients with alloimmune RSA.

The STAT Signaling Biosignature at the Single Cell Level Reveals Novel Insights in the Association of FOXP3+ T Regulatory Cells with Recurrent Spontaneous Abortions (RSA)

BackgroundTregs play an important role in the establishment and maintenance of immune tolerance during pregnancy and they are expanded early after embryo implantation. Decreased frequency and function of Tregs in women suffering from recurrent spontaneous abortions (RSA) has been reported in literature. Signal transducer and activator of transcription (STAT) proteins play a prominent role in the epigenetic control of Treg differentiation. Deficiency of STAT5 leads to loss of Foxp3+ T regulatory cells (Tregs) and inability to induce Tregs in vitro whereas constitutive activation of STAT5β enforces Foxp3 positive Treg development. Aberrant immune signaling via STATs is involved in disease pathobiology in numerous autoimmune conditions, therefore explored the STAT signaling biosignature of Tregs by single-cell phospho-specific flow cytometry in women with RSA before and after treatment with lymphocyte immunotherapy (LIT).Materials and methodsPeripheral blood mononuclear cells from 31 women with either RSA (n=23) or with infertility and ≥2 unsuccessful IVF (n=8) and 10 age-matched females without RSA who had at least one successful pregnancy, were obtained before and after LIT. Basal levels of phospho-STAT1, STAT3 and STAT5 and potentiated, i.e. target/stimuli, nodes (IFNα/pSTAT1, IFNα/pSTAT5, IL-6/pSTAT1, IL-6/pSTAT3 and IL-2/pSTAT5) were measured by phospho-specific flow cytometry. Treg signaling profiles were grouped by using unsupervised hierarchical clustering and correlated with clinical parameters by using χ2 or Fisher Exact tests. Associations of single signaling nodes with the aforementioned parameters were performed by Mann-Whitney and Wilcoxon signed-rank tests as appropriate.ResultsNo differences were observed between women with RSA and age matched women without RSA concerning the pretreatment and post treatment levels of Tregs. By contrast, clustering of pretreatment signaling nodes identified three signaling clusters (SC, figure 1a). SC#3 was enriched in women without RSA (p<0.001) and characterized by high potentiated responses of STATs, whereas women with RSA segregated in SC#1 and SC#2 which were characterized by blunted STAT responses in cytokine stimulation. Of note, all women with successful pregnancy after LIT (n=7) were clustered in SC#1 (p=0.028) which was characterized by high basal levels of pSTAT1 and pSTAT5 but weak potentiated responses of all nodes. Also, Tregs in women with a successful pregnancy after LIT displayed a significantly decreased response of pSTAT5 to IFNα (p= 0.05) and increased basal pSTAT5 levels (p=0.047), compared to women who failed LIT. Τhe post-treatment STAT signaling biosignature remained abnormal in women with RSA (p<0.001, figure 1b). However, compared to women with at least one miscarriage (n=23), women with infertility and unsuccessful IVFs displayed decreased post treatment basal levels of pSTAT1 (p=0.004) and pSTAT3 (p=0.01) and increased responses of pSTAT1 (p=0.047) and pSTAT3 (p=0.047) to IL6. Also, clustering the fold change of pre versus post-treatment STAT signaling profiles in Tregs revealed two signaling clusters. The #SC2 was marginally enriched in women with successful pregnancy after LIT (p=0.1) and was characterized by upregulation of all potentiated nodes after treatment (figure 1c).Collectively, we show for the first time that Tregs in women with RSA bear a pathological STAT biosignature which correlates with LIT outcome. Moreover, LIT can potentially induce a beneficial modulation of STAT signaling by upregulating Treg responsiveness to cytokines. Our work provides novel biological data on the mechanisms of immune tolerance in pregnancy and may point to therapies that can overcome pregnancy loss by modifying cell and cytokine responses. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Clinical effect of lymphocyte immunotherapy on patients with unexplained recurrent spontaneous abortion.

IntroductionLymphocyte immunotherapy (LIT) is believed to be a viable treatment for unexplained recurrent spontaneous abortion (URSA), but its effect remains controversial. This study aims to investigate the clinical effect of LIT in patients with URSA and clarify the factors that may influence the outcome of LIT.MethodsThis study included a total of 704 URSA patients, of which 444 patients accepted LIT treatment. URSA patients that did not accept LIT served as control group. Clinical characteristics were collected and analyzed between LIT and control group. The blocking antibody was tested before and after LIT. The outcome of LIT treatment was recorded. Logistic regression analysis was applied to evaluate the independent predictors of LIT success.ResultsAfter LIT treatment, 77.9% (346/444) of USRA patients turned to BA positive, and the conversion rate elevated with increased LIT (p < .001). LIT significantly improved the pregnancy rate and live birth rate in USRA patients (65.3% vs. 29.6%, p < .001; 80.3% vs. 50.6%, p < .001). Multivariate regression analysis suggested that younger maternal age and positive BA were independent predictors of LIT success.ConclusionLIT effectively induced the production of BA, and improved pregnancy rate and live birth rate in URSA patients. Our findings supported LIT as a beneficial treatment for URSA.

The Effects of LIT and MLR-Bf on Immune Biomarkers and Pregnancy Outcomes in Women With Previous Early Recurrent Miscarriage: A Retrospective Study.

Background: Immunological failure during pregnancy is considered one of the etiologies of recurrent miscarriage (RM). The decreased production of mixed lymphocyte reaction-blocking factors (MLR-Bf) may play a major role in this condition. Lymphocyte immunotherapy (LIT), which induces the production of MLR-Bf, has been used in treating RM patients since 1984. However, the effectiveness of LIT is currently being heatedly debated. In addition to that, possible changes to the maternal immune system upon induced MLR-Bf production by LIT remains unclear.Objectives: To explore the possible impacts that MLR-Bf may have on the expression of immune biomarkers and pregnancy outcomes, and deduce whether the prevention of miscarriages is possible with LIT or MLR-Bf in RM patients.Materials and Methods: Women with previous early RM (eRM) were enrolled in this retrospective study after they got pregnant again. LIT was implemented before pregnancy and during the first trimester. MLR-Bf and immune biomarkers were checked as the clinical routine. Patients were followed up until 12 gestational weeks. Levels of immune biomarkers and successful pregnancy rates were compared between MLR-Bf− group and MLR-Bf+ group stratified by LIT. Independent associations between LIT, or MLR-Bf, and miscarriage were estimated. All data management and analysis were conducted using SPSS 20.0.Results: A total of 1,038 patients, 497 MLR-Bf− (49 cases accepted LIT), and 541 MLR-Bf+(463 cases induced by LIT) were included in the study. Percentage of lymphocytes, the ratio of CD4+ T cells/lymphocytes, and levels of some rheumatoid biomarkers (anti-U1-nRNP, anti-SAA-52kd, and anti-CENOP B) were statistically higher in MLR-Bf+ group than in MLR-Bf− group among women without LIT. With LIT treatment the successful pregnancy rate was statistically higher in MLR-Bf+ group than in MLR-Bf− group (66.7% vs. 51.0%, P = 0.028) among women with LIT. Meanwhile, LIT was estimated to have an independent negative association with miscarriage.Conclusion: Upon LIT treament levels of immune biomarkers were different in women with and without MLR-Bf when stratified by whether they received LIT. Not MLR-Bf, but LIT, has an independent protective effect on miscarriage.

P80: Efficacy of lymphocyte immunotherapy in the treatment of recurrent pregnancy loss: A systematic review and meta‐analysis

P80: Efficacy of lymphocyte immunotherapy in the treatment of recurrent pregnancy loss: A systematic review and meta‐analysis