Targeting amino acid metabolism in cancer growth and anti-tumor immune response

Recent advances in amino acid metabolism have revealed that targeting amino acid metabolic enzymes in cancer therapy is a promising strategy for the development of novel therapeutic agents. There are currently several drugs in clinical trials that specifically target amino acid metabolic pathways in tumor cells. In the context of the tumor microenvironment,however,tumor cells form metabolic relationships with immune cells,and they oftencompete for common nutrients. Many tumors evolved to escape immune surveillance by taking advantage of their metabolic flexibility and redirecting nutrients for their own advantage. This review outlines the most recent advances in targeting amino acid metabolic pathways in cancer therapy while giving consideration to the impact these pathways may have on the anti-tumor immune response.

Immunization with autologous T cells enhances in vivo anti-tumor immune responses accompanied by up-regulation of GADD45β

Immunization with inactivated autoreactive T cells may induce idiotype anti-idiotypic reactions to deplete autoreactive T cells, which are involved in autoimmune diseases. However, it is unknown whether attenuated activated healthy autologous T-cell immunization could increase anti-tumor immune responses. To this end, C57B1/6 mice were immunized with attenuated activated autologous T cells. The splenocytes from immunized mice showed a higher proliferative ability than that from naive mice. The special phenotype analysis showed that there were more CD8+ T cells and CD62L+ T cells in immunized mice after 24 h of culture with 10% fetal calf serum complete medium in vitro (P〈0.01). These results demonstrated that this immunization may activate T cells in vivo. Furthermore, the splenocytes from immunized mice revealed resistance to activation-induced cell death (AICD) in vitro. To further study the relative genes that are responsible for the higher proliferation and resistance to AICD, the expression of Fas/Fas ligand (FasL) and GADD4513 was measured by real-time PCR. The results indicated that GADD45β transcription was higher in the splenocytes from immunized mice than that in the naive mice. In addition, the Fas expression showed a parallel higher, but FasL did not change obviously. To investigate the biologic functions induced by immunization in vivo, a tumor model was established by EL-4 tumor cell inoculation in C57/B1 mice. Mice receiving autologous T-cell immunization had significantly inhibited tumor growth in vivo (P〈0.01). This study implicated that immunization with attenuated activated autologous T cells enhances anti-tumor immune responses that participate in tumor growth inhibition.

Anti-tumor Immune Response Mediated by Newcastle Disease Virus HN Gene

Hemagglutinin-neuramidinase（HN） is one of the most important surface structure proteins of the Newcastle disease virus（NDV）. HN not only mediates receptor recognition but also possesses neuraminidase（NA） activity, which gives it the ability to cleave a component of those receptors, NAcneu. Previous studies have demonstrated that HN has interesting anti-neoplastic and immune-stimulating properties in mammalian species, including humans. To explore the application of the HN gene in cancer gene therapy, we constructed a Lewis lung carcinoma（LLC） solid tumor model using C57BL/6 mice. Mice were injected intratumorally with the recombinant adenovirus expressing HN gene（Ad-HN）, and the effect of HN was explored by natural killer cell activity assay, cytotoxic lymphocyte activity assay, T cell subtype evaluation, and Th1/Th2 cytokines analysis. The results demonstrate that HN not only can elicit clonal expansion of both CD4＋ and CD8＋ T cell populations and cytotoxic T lymphocyte（CTL） and killer cell response, but also skews the immune response toward Th1. Thus, vaccination with Ad-HN may be a potential strategy for cancer gene therapy.

Effect of postoperative early enteral nutrition support on anti-tumor immune response and inflammatory response process in the elderly patients with esophageal cancer

Objective: To explore the effect of postoperative early enteral nutrition support on anti-tumor immune response and inflammatory response process in the elderly patients with esophageal cancer. Methods: A total of 110 cases of elderly patients with esophageal cancer who underwent radical operation in this hospital between January 2015 and December 2017 were divided into the parenteral nutrition group (n=57) who received parenteral nutrition support and the enteral nutrition group (n=53) who received enteral nutrition support according to the postoperative nutrition intervention methods. The differences in anti-tumor immune response and inflammatory response degree were compared between the two groups of patients immediately after the patients returned to the ward (T0), 48h after nutritional support (T1) and 72h after nutritional support (T2). Results: At T0, there was no statistically significant difference in the contents of Th1/Th2 immune response and Th17/Treg immune response indexes as well as inflammatory mediators in serum between the two groups of patients. At T1 and T2, serum Th1 cytokines IFN-γ and IL-12 contents of enteral nutrition group were higher than those of parenteral nutrition group whereas Th2 cytokines IL-4 and IL-13 contents were lower than those of parenteral nutrition group;serum contents of Th17 cytokines IL-6 and IL-17 as well as Treg cytokines TGF-β and IL-10 were lower than those of parenteral nutrition group;serum inflammatory mediators hs-CRP, PGE and HMGB1 contents were lower than those of parenteral nutrition group. Conclusion: Postoperative early enteral nutrition support can effectively regulate the Th1/Th2 and Th17/Treg immune response balance and inhibit the systemic inflammatory response in elderly patients with esophageal cancer.

Effects of different chemotherapy regimens before radical operation for lung cancer on cancer cell growth and anti-tumor immune response

Objective:To investigate the effects of different chemotherapy regimens before radical operation for lung cancer on cancer cell growth and antitumor immune response.Methods: A total of 180 patients with primary lung cancer who underwent surgery in this hospital between February 2013 and August 2017 were divided into the cisplatin group (n=93) who received cisplatin & paclitaxel chemotherapy and the lobaplatin group (n=87) who received lobaplatin & paclitaxel chemotherapy according to different preoperative neoadjuvant chemotherapy regimens. The differences among the expression of proliferation genes and apoptosis genes in tumor tissues as well as the contents of Th1/Th2 cytokines in serum were compared between the two groups.Results: Proliferation genes DDX17, GPx1, MACC1, RACK1 and SIRT1 mRNA expression levels in tumor tissue of lobaplatin group were lower than those of cisplatin group whereas LRRC3B mRNA expression level was higher than that of cisplatin group;apoptosis gene Fas, FasL and Caspase-3 mRNA expression levels were higher than those of cisplatin group whereas Survivin and Bcl-2 mRNA expression levels were lower than those of cisplatin group;serum Th1 cytokines IFN-γ and IL-12 contents were higher than those of cisplatin group whereas Th2 cytokines IL-5 and IL-10 contents were lower than those of cisplatin group.Conclusion: Lobaplatin chemotherapy before radical operation for lung cancer is more effective than cisplatin chemotherapy to inhibit the proliferation activity and enhance the apoptosis activity of lung cancer cells and optimize the anti-tumor immune response.

Effect of dietary fiber enteral nutrition on inflammatory response and anti-tumor immune response after colorectal cancer surgery

Objective: To study the effect of dietary fiber enteral nutrition on inflammatory response and anti-tumor immune response after colorectal cancer surgery. Methods: A total of 162 patients who underwent radical operation for colorectal cancer in West China Hospital, Sichuan University between March 2015 and August 2017 were selected as the research subjects and randomly divided into the dietary fiber group who accepted dietary fiber enteral nutrition intervention and the control group who accepted conventional nutrition intervention. The number of intestinal flora was determined before surgery and at the first defecation after surgery;the inflammatory response indexes and immune response indexes were measured before surgery as well as 3 d and 5 d after surgery. Results: At the first defecation after surgery, the bifidobacteria, lactobacillus, Escherichia coli and enterococcus contents in the feces of both groups were lower than those before surgery, and the bifidobacterium and lactobacillus contents in feces of dietary fiber group were higher than those of control group whereas Escherichia coli and enterococcus contents were lower than those of control group;3 d and 5 d after surgery, NF-κB, IL-1, TNF-α and IL-8 contents in serum as well as Treg, Th17, Th22, TGF-β, IL-17 and IL-22 contents in peripheral blood of both groups of patients were significantly higher than those before surgery, and NF-κB, IL-1, TNF-α and IL-8 contents in serum as well as Treg, Th17, Th22, TGF-β, IL-17 and IL-22 contents in peripheral blood of dietary fiber group were lower than those of control group. Conclusion: The dietary fiber enteral nutrition intervention after colorectal cancer surgery can inhibit the inflammatory response and improve the anti-tumor immune response.

Effect of negative emotion on tumor load and anti-tumor immune response in bladder cancer infusion chemotherapyn

Objective: To study the effect of negative emotion on tumor load and anti-tumor immune response in bladder cancer infusion chemotherapy. Methods: Patients with advanced bladder cancer who received bladder infusion chemotherapy in the First People's Hospital of Ziyang between May 2014 and December 2016 were selected and divided into the control group without negative emotions, the anxiety group with anxiety, the depression group with depression and the anxiety depression group with both anxiety and depression according to the assessment results of HAMA scale and HAMD scale. The contents of tumor markers in serum, the contents of immune cells in peripheral blood and the expression of apoptosis genes in urine were detected during chemotherapy. Results: DKK-1, DKK-3, OPN and CYFRA21-1 contents in serum as well as PD-1+CD4+T cell, PD-1+CD8+T cell and B7-H1+CD11c+DC cell contents in peripheral blood of anxiety group, depression group and anxiety depression group were significantly higher than those of control group while DAP2IP, Fas, FasL, Caspase-8 and PTEN protein expression in urine were significantly lower than those of control group, and DKK-1, DKK-3, OPN and CYFRA21-1 contents in serum as well as PD-1+CD4+T cell, PD-1+CD8+T cell and B7-H1+CD11c+DC cell contents in peripheral blood of anxiety depression group were significantly higher than those of anxiety group and depression group while DAP2IP, Fas, FasL, Caspase-8 and PTEN protein expression in urine were significantly lower than those of anxiety group and depression group. Conclusion: The anxiety and depression in bladder cancer infusion chemotherapy can suppress the expression of apoptosis genes and the anti-tumor immune response to lead to more vigorous proliferation of cancer cells.

Changes of serum endocrine hormone levels in patients with cancer-related fatigue and their correlation with anti-tumor immune response and tumor load

Objective: To study the changes of serum endocrine hormone levels in patients with cancer-related fatigue (CRF) and their correlation with anti-tumor immune response and tumor load. Methods: A total of 137 patients who were diagnosed with primary lung cancer in West China Hospital, Sichuan University between June 2014 and November 2016 were selected and then divided into CRF group and control group according to their self-reported symptoms, serum was collected to determine the levels of endocrine hormones and tumor markers, and peripheral blood was collected to detect the levels of immune cells. Results: Serum ACTH and TSH levels of CRF group were significantly higher than those of control group while Cor, FT3 and FT4 levels were significantly lower than those of control group;peripheral blood CD11b+CD15-CD33+CD14+M-MDSC, CD11b+CD15-CD33+CD14-G-MDSC, CD4+CD25+CD127lowTreg and CD19+CD5+CD1d+Breg levels as well as serum CEA, Cyfra21-1, SCC-Ag, HE4, GDF-15 and PCNA levels of CRF group were significantly higher than those of control group, positively correlated with serum ACTH and TSH levels, and negatively correlated with Cor, FT3 and FT4 levels. Conclusion: The changes of thyroid hormone and adrenal cortical hormone levels in patients with cancer-related fatigue are closely related to the inhibited anti-tumor immune response and increased tumor load.

Effect of enteral immunonutrition after radical surgery for esophageal carcinoma on anti-tumor immune response and intestinal mucosal barrier function

Objective:To study the effect of enteral immunonutrition after radical surgery for esophageal carcinoma on anti-tumor immune response and intestinal mucosal barrier function.Methods:A total of102 patients who received radical surgery for esophageal carcinoma in our hospital between May 2013 and December 2016 were selected and randomly divided into observation group and control group who received postoperative enteral immunonutrition and routine enteral nutrition respectively. 1 d before operation as well as 1 d and 7 d after operation, peripheral blood immune cell marker expression and serum intestinal mucosal barrier injury marker levels were detected.Results:1 d after operation, peripheral blood T-bet, NKG2D, NKp30, NKp44 and NKp46 fluorescence intensity of both groups of patients were significantly lower than those 1d before operation while peripheral blood GATA-3 and Foxp3 fluorescence intensity as well as serum DAO, Occludin, ZO-1 and claudin-1 levels were significantly higher than those 1d before operation;peripheral blood T-bet, NKG2D, NKp30, NKp44 and NKp46 fluorescence intensity of observation group 7 d after operation were significantly higher than those 1 d after operation while peripheral blood GATA-3 and Foxp3 fluorescence intensity as well as serum DAO, Occludin, ZO-1 and claudin-1 levels were significantly lower than those 1 d after operation;peripheral blood T-bet, GATA-3, Foxp3, NKG2D, NKp30, NKp44 and NKp46 fluorescence intensity of control group 7 d after operation were not significant different from those 1 d after operation, and serum DAO, Occludin, ZO-1 and claudin-1 levels were significantly lower than those 1d after operation.Conclusion: Enteral immunonutrition after radical surgery for esophageal carcinoma can enhance the anti-tumor immune response and improve the intestinal mucosal barrier function.

Effects of Rosa roxburghii&edible fungus fermentation broth on immune response and gut microbiota in immunosuppressed mice

With the rise of probiotics fermentation in food industry,fermented foods have attracted worldwide attention.In this study,protective effects of Rosa roxburghii&edible fungus fermentation broth(REFB)on immune function and gut health in Cyclophosphamide induced immunosuppressed mice were investigated.Results showed that REFB could improve the immune organ index,and promote the proliferation and differentiation of splenic T lymphocytes.In addition,it attenuated intestinal mucosal damage and improved intestinal cellular immunity.REFB administration also up-regulated the expression of IL-4,INF-γ,TNF-α,T-bet and GATA-3 mRNA in small intestine.Furthermore,administration of REFB modulated gut microbiota composition and increased the relative abundance of beneficial genus,such as Bacteroides.It also increased the production of fecal short-chain fatty acids.These indicate that REFB has the potential to improve immunity,alleviate intestinal injury and regulate gut microbiota in immunosuppressed mice.

Effect of dietary supplement of inactivated Lactobacillus plantarum Ep-M 17 on growth performance,immune response,disease resistance,and intestinal microbiota in Penaeus vannamei

Our previous study found that feeding with Lactobacillus plantarum Ep-M17 could effectively affect the growth performance,immune response,and gut microbiota of Penaeus vannamei.However,high temperature and pressure during feed pelletizing is the main problem that can lead to a decrease in the activity of probiotics or cause their inactivation.Further investigation needs to investigate whether inactivated Ep-M17 can exert similar effects as live Ep-M17.Therefore,we evaluated the effects of inactivated L.plantarum Ep-M17 on growth performance,immune response,disease resistance,and gut microbiota in P.vannamei.Results show that adding inactivated Ep-M17 to the feed also promoted body weight gain and increased relative immune protection in shrimp.Also,histological examination revealed that the administration of inactivated Ep-M17 led to improvements in the density and distribution of microvilli in the intestines and enhancements in the abundance of B and R cells in the hepatopancreas.Additionally,the inactivated Ep-M17 supplementation resulted in increased activity levels of nutrient immune-related enzymes in both the shrimp hepatopancreas and intestines.Moreover,it stimulated the expression of Lvlec,PEN-3a,Crustin,LGBP,Lysozyme,and proPo genes in both the hepatopancreas and intestines.Furthermore,the inactivated Ep-M17 also increased bacterial diversity in the gut of shrimp and promoted the abundance of specific flora,facilitating the host organism’s metabolism and immunity to improve the disease resistance of shrimp.Therefore,supplementation of inactivated L.plantarum Ep-M17 in shrimp diets can exert similar effects as live L.plantarum Ep-M17 effectively improving growth performance,gut microbiota,immune response,and disease resistance in P.vannamei.

NIR-triggered on-site NO/ROS/RNS nanoreactor:Cascade-amplified photodynamic/photothermal therapy with local and systemic immune responses activation

Photothermal and photodynamic therapies(PTT/PDT)hold promise for localized tumor treatment,yet their full potential is hampered by limitations such as the hypoxic tumor microenvironment and inadequate systemic immune activation.Addressing these challenges,we present a novel near-infrared(NIR)-triggered RNS nanoreactor(PBNO-Ce6)to amplify the photodynamic and photothermal therapy efficacy against triple-negative breast cancer(TNBC).The designed PBNOCe6 combines sodium nitroprusside-doped Prussian Blue nanoparticles with Chlorin e6 to enable on-site RNS production through NIR-induced concurrent NO release and ROS generation.This not only enhances tumor cell eradication but also potentiates local and systemic antitumor immune responses,protecting mice from tumor rechallenge.Our in vivo evaluations revealed that treatment with PBNO-Ce6 leads to a remarkable 2.7-fold increase in cytotoxic T lymphocytes and a 62%decrease in regulatory T cells in comparison to the control PB-Ce6(Prussian Blue nanoparticles loaded with Chlorin e6),marking a substantial improvement over traditional PTT/PDT.As such,the PBNO-Ce6 nanoreactor represents a transformative approach for improving outcomes in TNBC and potentially other malignancies affected by similar barriers.

Immune response to inactivated bacterial vector carrying the recombinant K39 antigen of Leishmania infantum in mice

Objective:To evaluate the immunological response elicited by an inactivated bacterial vector carrying the K39 antigen of Leishmania infantum,and a purified antigen.Methods:Mice were subjected to the following treatments:(1)Purified recombinant K39(rK39)protein at a 20μg dose with complete Freund’s adjuvant;(2)Inactivated Escherichia coli(BL21 DE3)carrying the K39 protein at an equivalent total protein content of 200μg;(3)Inactivated bacteria lacking the K39 protein;(4)Non-immunized control animals.Serological monitoring was performed.All groups were challenged by intraperitoneal injection of 10^(7) Leishmania infantum promastigotes.After euthanasia,the liver and spleen were collected to analyze the levels of TNF,IFN-γ,IL-12,IL-4,and IL-10.Results:Mice immunized with purified rK39 or the inactivated bacterial vector carrying the K39 antigen of Leishmania infantum showed a long-lasting immune response with high levels of polyclonal antibodies specifically recognizing the recombinant proteins.The IgG1 subclass was the predominant immunoglobulin;however,the induction of IgG2a and the profile of cytokines produced were indicative of the induction of a mixed-type response.Conclusions:The inactivated bacterial vector carrying the K39 antigen,as well as the purified antigen can induce a long-lasting immune response in immunized mice,predominantly favouring a Th2 profile response.

Acupoint catgut-embedding therapy ameliorates DNCB-induced atopic dermatitis in BALB/c mice by regulating Th2 type immune response and reducing infiltration of CD4^(+)and CD8^(+)cells

Background:This study aimed to assess how acupoint catgut-embedding therapy influences Th2-type immune response and the infiltration of CD4^(+)and CD8^(+)cells in DNCB-induced atopic dermatitis in BALB/c mice.It also conducted an initial examination of the underlying molecular mechanisms.Methods:Seventy-two mice were randomly divided into four groups:normal control,DNCB-induced atopic dermatitis model(AD),AD with acupoint catgut-embedding treatment(ADA),and AD with sham-acupoint catgut-embedding treatment.After DNCB challenge to induce AD,the ADA group received acupoint catgut-embedding therapy treatment at Zusanli(ST 36)and Quchi(LI 11)acupoints every other week from day 8.Mice in the AD with sham-acupoint catgut-embedding treatment group underwent the same procedure as the ADA group but without catgut implantation.Severity was assessed using SCORAD on treatment days 1,10,and 20.On day 18,nine mice per group were euthanized,and the remaining on day 28.Histopathological changes were observed using hematoxylin-eosin and immunohistochemistry staining.TNF-α,IL-4,IL-6,and IL-13 levels were analyzed by ELISA,and GATA3 and STAT6 protein levels by western blot.Results:After 20 days of acupoint catgut-embedding therapy treatment,mice showed reduced dermatitis scores compared to DNCB-induced AD-like mice.Significant decreases occurred in serum IL-4,IL-6,IL-13,and TNF-αlevels.Skin analysis revealed marked reductions in CD4^(+)and CD8^(+)cell infiltration,as well as GATA3 and STAT6 protein levels.Conclusion:Acupoint catgut-embedding therapy may effectively alleviate atopic dermatitis by suppressing Th2 immune responses via the STAT6-GATA3 pathway and reducing CD4^(+)and CD8^(+)T cell infiltration in skin lesions.

Impact of minimally invasive surgery on immune function and stress response in gastric cancer patients

BACKGROUND Gastric cancer remains a leading cause of cancer-related mortality globally.Traditional open surgery for gastric cancer is often associated with significant morbidity and prolonged recovery.AIM To evaluate the effectiveness of laparoscopic minimally invasive surgery as an alternative to traditional open surgery for gastric cancer,focusing on its potential to reduce trauma,accelerate recovery,and achieve comparable oncological out-comes.METHODS This study retrospectively analyzed 203 patients with gastric cancer who underwent surgery at the Shanghai Health Medical College Affiliated Chongming Hospital from January 2020 to December 2023.The patients were divided into two groups:Minimally invasive surgery group(n=102),who underwent laparoscopic gastrectomy,and open surgery group(n=101),who underwent traditional open gastrectomy.We compared surgical indicators(surgical incision size,intraop-erative blood loss,surgical duration,and number of lymph nodes dissected),recovery parameters(time to first flatus,time to start eating,time to ambulation,and length of hospital stay),immune function(levels of IgA,IgG,and IgM),intestinal barrier function(levels of D-lactic acid and diamine oxidase),and stress response(levels of C-reactive protein,interleukin-6,and procalcitonin).RESULTS The minimally invasive surgery group demonstrated significantly better outcomes in terms of surgical indicators,including smaller incisions,less blood loss,shorter surgery time,and more lymph nodes dissected(P<0.05 for all).Recovery was also faster in the minimally invasive surgery group,with earlier return of bowel function,earlier initiation of diet,quicker mobilization,and shorter hospital stays(P<0.05 for all).Furthermore,patients in the minimally invasive surgery group had better preserved immune function,superior intestinal barrier function,and a less pronounced stress response postoperatively(P<0.05 for all).CONCLUSION Laparoscopic minimally invasive surgery for gastric cancer not only provides superior surgical indicators and faster recovery but also offers advantages in preserving immune function,protecting intestinal barrier function,and mitigating the stress response compared to traditional open surgery.These findings support the broader adoption of laparoscopic techniques in the management of gastric cancer.

Probiotics: Shaping the gut immunological responses

Probiotics are live microorganisms exerting beneficial effects on the host’s health when administered in adequate amounts.Among the most popular and adequately studied probiotics are bacteria from the families Lactobacillaceae,Bifidobacteriaceae and yeasts.Most of them have been shown,both in vitro and in vivo studies of intestinal inflammation models,to provide favorable results by means of improving the gut microbiota composition,promoting the wound healing process and shaping the immunological responses.Chronic intestinal conditions,such as inflammatory bowel diseases(IBD),are characterized by an imbalance in microbiota composition,with decreased diversity,and by relapsing and persisting inflammation,which may lead to mucosal damage.Although the results of the clinical studies investigating the effect of probiotics on patients with IBD are still controversial,it is without doubt that these microorganisms and their metabolites,now named postbiotics,have a positive influence on both the host’s microbiota and the immune system,and ultimately alter the topical tissue microenvironment.This influence is achieved through three axes:(1)By dis-placement of potential pathogens via competitive exclusion;(2)by offering protection to the host through the secretion of various defensive mediators;and(3)by supplying the host with essential nutrients.We will analyze and discuss almost all the in vitro and in vivo studies of the past 2 years dealing with the possible favorable effects of certain probiotic genus on gut immunological responses,highlighting which species are the most beneficial against intestinal inflammation.

OsbZIP53 Negatively Regulates Immunity Response by Involving in Reactive Oxygen Species and Salicylic Acid Metabolism in Rice

The basic region/leucine zipper(bZIP)transcription factors play important roles in plant development and responses to abiotic and biotic stresses.OsbZIP53 regulates resistance to Magnaporthe oryzae in rice by analyzing APIP5-RNAi transgenic plants.To further investigate the biological functions of OsbZIP53,we generated osbzip53 mutants using CRISPR/Cas9 editing and also constructed OsbZIP53 over-expression transgenic plants.Comprehensive analysis of phenotypical,physiological,and transcriptional data showed that knocking-out OsbZIP53 not only improved disease resistance by inducing a hypersensitivity response in plants,but also regulated the immune response through the salicylic acid pathway.Specifically,disrupting OsbZIP53 increased H2O2 accumulation by promoting reactive oxygen species generation through up-regulation of several respiratory burst oxidase homologs(Osrboh genes)and weakened H2O2 degradation by directly targeting OsMYBS1.In addition,the growth of osbzip53 mutants was seriously impaired,while OsbZIP53 over-expression lines displayed a similar phenotype to the wild type,suggesting that OsbZIP53 has a balancing effect on rice immune response and growth.

Immune Response of the Ridgetail White Prawn Exopalaemon carinicauda After Exposure to the Dinoflagellate Prorocentrum minimum

The dinoflagellate Prorocentrum minimum is known to affect the normal physiological function of Exopalaemon carinicauda by inducing oxidative stress,apoptosis,and cellular injury.To study the effects of P.minimum on the immune defense system of shrimp,E.carinicauda were exposed to 5×10^(3)cells mL^(-1)and 5×10^(4)cells mL^(-1)of P.minimum for 336 h in treatment groups,while E.carinicauda cultured in filtered seawater was employed as control.The total hemocyte counts(THC),hemocyanin concentration(HEM),and the activity of alkaline phosphatase(AKP)in hemolymph serum,as well as expressions of six immunity-related genes in hemocytes,hepatopancreases and gills were determined.The exposure of P.minimum significantly reduced the THC,HEM concentration and AKP activity in hemolymph serum.Immunity-related genes expressed differently in hemocytes,hepatopancreases and gills.Compared with the control group,the expressions of Crustin and pro PO in hemocytes were significantly up-regulated in the treatment groups,while the up-regulated expressions of LGBP,Lysozyme and Serpin were only found in the group exposed to 5×10^(4)cells mL^(-1)of P.minimum.In the gills of E.carinicauda exposed to P.minimum,the down-regulation of ALF,proPO and Serpin,up-regulation of LGBP and Lysozyme,as well as unaffected Crustin were observed.In hepatopancreases,the up-regulated expressions of LGBP,Crustin,Lysozyme,Serpin and proPO(only in 5×10^(3)cells mL^(-1)of P.minimum group)were found in the treatment groups.After exposure to P.minimum for 336 h,shrimps were injected with Vibrio parahaemolyticus and WSSV.The results showed that the mortality rates of shrimp in the treatment groups were significantly increased with a dose-dependent effect,which suggests that exposure to P.minimum may reduce the immunity of E.carinicauda.The research indicates that hemocytes and hepatopancreases play important roles in protecting the shrimp immune response to harmful algae,while the protection effect of hemolymph serum and gills may be suppressed.Since the exposure to P.minimum depressed the immunity of E.carinicauda,further studies are needed to confirm whether the presence of the algae will affect the susceptibility of shrimp to pathogens.

Comparison of Pfizer/BioNTech BNT162b2 COVID-19 (Comirnaty) Vaccination Effects in Aged Adults with Special Consideration for the Effectiveness of the Three-Color iSpot in Assessing Immune Response

Background: The roll-out of vaccines against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was certainly among the fastest in medical history. Vaccination campaigns around the world began a year after the outbreak in 2019. When Austria started vaccinating the population in 2020, we took the opportunity to collect data from the first sets of patients receiving the vaccine in our study region of East Tyrol. Purpose: Many studies have been conducted examining the immunogenicity of the new vaccines using classic serological test methods in combination with an IFN-γ ELISpot. Undeniable disadvantages of using IFN-γ to characterize the status of the cellular immunity are that 1) being an acute phase cytokine, IFN-γ loses signal strength in the long run and 2) IFN-γ does not provide information about the involvement of T helper 2 (Th2) cells in the immune process. This implies that it can affect false negative data about the cell-mediated immune status. Method: Therefore, in addition to a chemiluminescent immunoassay and the enzymatic IFN-γ ELISpot, this study included a fluorescent ELISpot assay using precoated human SARS-CoV-2-specific IFN-γ/IL-2/IL-5 ELISpot kits to show a more holistic overview on the involvement of T helper 1 (Th1) cells as signal senders of IL-2 and Th2 cells as senders of IL-5. Results and Conclusion: Our study confirms good immunogenicity of Pfizer/BioNTech BNT162b2 COVID-19 (Comirnaty) with strong Th1 and vanishingly small Th2 participation. The fluorescent three color iSpot can improve the diagnostic results’ significance for the individual, especially when the infection has been longer in the past and the IFN-γ signal diminishes.

CRISPR/Cas9-mediated knockout of SLC15A4 gene involved in the immune response in bovine rumen epithelial cells

The objective of this study was to determine the role of SLC15A4 in the muramyl dipeptide(MDP)-mediated inflammatory response of bovine rumen epithelial cells(BRECs).First,changes in the m RNA expression of proinflammatory factor genes in BRECs following 10μg m L^(–1)MDP treatments were examined.RT-q PCR results showed that the expression of pro-inflammatory factor(IL-1β,IL-6,and TNF-α)m RNAs were significantly increased under MDP stimulation(P<0.001).Moreover,SLC15A4-Knockout(SLC15A4-KO)cells were obtained through lentivirus packaging,transfection,screening,and cell monoclonal culture.In order to gain further insight into the potential function of SLC15A4,we utilized transcriptome data,which revealed a change in the genes between WT-BRECs and SLC15A4-KO.Five down-regulated pro-inflammatory genes and 13 down-regulated chemokine genes related to the inflammatory response were identified.Meanwhile,the down-regulated genes were mostly enriched in the nuclear factorκB(NF-κB)and mitogen-activated protein kinase(MAPK)signaling pathways.The results of RT-q PCR also verified these detected changes.To further determine the mechanism of how WT and SLC15A4-KO BRECs are involved in inflammatory responses,we investigated the inflammatory responses of cells exposed to MDP.WT-BRECs and SLC15A4-KO were treated with a culture medium containing 10μg m L^(–1)MDP,in comparison to a control without MDP.Our results show that SLC15A4-KO BRECs had reduced the expression of genes(IL-6,TNF-α,CXCL2,CXCL3,CXCL9,and CCL2)and proteins(p-p65 and p-p44/42)from the MDP-mediated inflammatory response compared to WT-BRECs(P<0.05).In this experiment,CRISPR-Cas9 was used to KO the di/tripeptide transporter SLC15A4,and its role was confirmed via the MDP-induced inflammatory response in BRECs.This work will provide a theoretical basis for studying the pro-inflammatory mechanism of MDP and its application in the prevention and treatment of subacute rumen acidosis in dairy cows.