Lymphocyte Stimulation Test Research Articles

Evaluation of the drug-induced lymphocyte stimulation test for diagnosing mesalazine allergy.

Background/AimsMesalazine is an effective drug for treating ulcerative colitis (UC), but causes allergic symptoms in a few cases. Therefore, the objective of this study was to evaluate the usefulness of the drug-induced lymphocyte stimulation test (DLST) for the diagnosis of mesalazine allergy.MethodsPatients with UC treated with mesalazine with or without a history of associated adverse events (AEs) were enrolled at Kyorin University Hospital from July 2016 to April 2017.ResultsThe DLST was performed in 104 patients with UC, of which 24 had a history of AEs due to mesalazine treatment. The control value of DLST was 337.4±296.3 counts per minute (cpm) in the AE+ group and 408.0±371.9 cpm in the AE− group. The measured value of DLST was 578.8±424.7 cpm in the AE+ group and 476.5±471.8 cpm in the AE− group. The stimulation index (SI) was 243.9%±291.1% in the AE+ group and 119.8%±53.0% in the AE− group. The SI value and DLST positivity were significantly higher in the AE+ group than in the AE− group (P=0.030 and P=0.029, respectively). The test sensitivity and specificity were 0.240 and 0.805, respectively, and the false-positive and false-negative rate was 0.195 and 0.760, respectively.ConclusionsThe DLST for mesalazine showed low sensitivity and high specificity, suggesting that it may be useful for the definitive diagnosis of allergy to mesalazine.

USEFULNESS OF LYMPHOCYTE STIMULATION TEST AGAINST PIGEON SERUM IN MANAGEMENT OF PATIENTS WITH CHRONIC HYPERSENSITIVITY PNEUMONITIS

RespirologyVolume 22, Issue S3 p. 256-256 APSR Poster Abstracts USEFULNESS OF LYMPHOCYTE STIMULATION TEST AGAINST PIGEON SERUM IN MANAGEMENT OF PATIENTS WITH CHRONIC HYPERSENSITIVITY PNEUMONITIS First published: 23 November 2017 https://doi.org/10.1111/resp.13207_453Citations: 1Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article.Citing Literature Volume22, IssueS3Supplement: 22nd Congress of the Asian Pacific Society of Respirology, International Convention Centre, Sydney, Australia, 23–26 November 2017November 2017Pages 256-256 RelatedInformation

Seasonal Influenza Vaccine-induced Pneumonitis Presenting with Multiple Pulmonary Nodules

A 39-year-old woman received a seasonal influenza vaccine in November 2015 and subsequently experienced malaise, low-grade fever, and chest discomfort. A chest X-ray performed 2 weeks after vaccination showed multiple nodular shadows in both lungs and ground-glass shadows in both lower lung fields. Her bronchoalveolar lavage fluid contained an unusually high number of lymphocytes, and a drug-induced lymphocyte stimulation test for seasonal influenza vaccine was positive. Transbronchial lung biopsy revealed the presence of granulomatous inflammation. Thereafter her abnormal chest shadow spontaneously improved. Based on these findings, the patient was diagnosed with drug-induced pneumonitis due to an influenza vaccine.

Effect of probiotic, prebiotic and synbiotic on faecal microbial count and cell-mediated immunity in crossbred calves

A study was conducted to evaluate the impact of probiotic, prebiotic and synbiotic as feed additives on faecal counts of coliform and E. coli along with cell mediated immune response in crossbred calves. For this purpose fifteen days old, 24 crossbred calves were divided into 4 groups of 6 calves each. The calves were assigned to different dietary treatments viz T0(control group) -basal diet, T1 (probiotic group) - basal diet + Lactobacillus acidophilus @ 1 g/calf/day (2 x 1010 cfu/ g), T2 (prebiotic group)- basal diet + Mannan oligosaccharide (MOS) @ 4 g per calf/day and T3 (synbiotic group) basal diet +Lactobacillus acidophilus @ 0.5 gm (2 x 1010 cfu/ g) + 2 g MOS per calf/day respectively for a period of 90 days. Faecal samples for coliform and E. coli count were taken at 15 days interval. Blood samples were taken at 15 days interval for lymphocyte count. Blood samples were taken at 45th and 90th-day for lymphocyte stimulation test (LST) in EDTA coated vials. Skin fold test was done on 90th day of experimental study. The results of study showed that there was a significant reduction in faecal shedding of coliform and E. coli in T1, T2 and T3 groups indicating lesser proliferation of pathogenic bacteria in the gut. The treatment groups T2 and T3 showed a significant increase in lymphocyte count whereas all the treated groups had significantly higher values of the lymphocytic index on 45th and 90th day post supplementation indicating improved cell mediated immunity. Increased skin fold thickness was observed post 48 hours of phytohaemagglutinin injection in T2 and T3 groups. It may, therefore, be concluded that supplementation of prebiotic and synbiotic is advantageous in attaining gut conditioning and enhanced immunity which may be helpful in preventing calf scours in crossbred calves.

Second-line therapy for Helicobacter pylori eradication causing antibiotic-associated hemorrhagic colitis

ObjectiveHelicobacter pylori (H. pylori) eradication rarely develops into antibiotic-associated hemorrhagic colitis (AAHC), in which the etiology of colitis remains unclear. We herein report a rare case of AAHC caused by second-line therapy for H. pylori eradication.ResultsA 65-year-old female was administered second-line therapy for H. pylori composed of 1500 mg of amoxicillin, 500 mg of metronidazole and 40 mg of vonoprazan for 7 days because of first-line therapy failure. A day after completing second-line therapy, she complained of abdominal pain and hematochezia. Colonoscopy revealed a hemorrhage and edematous mucosa with no transparent vascular pattern in the transverse colon. A bacterial culture detected Klebsiella oxytoca (K. oxytoca), but no other pathogenic bacteria. A drug-induced lymphocyte stimulation test (DLST) showed positive reactions for both amoxicillin and metronidazole. According to these findings, the patient was diagnosed with AAHC. Bowel rest for 6 days relieved her abdominal pain and hematochezia.ConclusionsThe present case developed AAHC caused by second-line therapy for H. pylori eradication. The pathogenesis is considered to be associated with microbial substitution as well as a delayed-type allergy to antibiotics, suggesting that AAHC is a potential adverse event of second-line therapy for H. pylori eradication.

A case of acetaminophen-induced acute tubulointerstitial nephritis in adult.

We report a case of allergic acute tubulointerstitial nephritis (TIN) induced by acetaminophen in a 48-year-old Japanese man with no past medical history. Two days after receiving the non-steroidal anti-inflammatory drug (NSAID) loxoprofen for left shoulder pain, he developed cold symptoms such as fever and sore throat. He then took a 300mg dose of acetaminophen three times a day and a 100mg dose of minocycline hydrochloride twice a day for 7days. Because there was no improvement in his symptoms, he consulted a local clinic again, where blood tests revealed renal insufficiency, and he was, then, referred to our hospital for evaluation of kidney function. Renal biopsy revealed acute TIN, and Ga-67 scintigraphy showed diffuse uptake in bilateral kidneys. A drug-induced lymphocyte stimulation test (DLST) was positive for acetaminophen and negative for loxoprofen and minocycline. Based on these findings, we made a diagnosis of acetaminophen-induced TIN. We treated the patient with three courses of semi-pulse steroid therapy, after which his fever went down, and his serum creatinine level recovered from 2.09 to 1.43mg/dL. Although we medical doctors think that therapeutic dose of acetaminophen retains high safety, it is important to keep in mind that acetaminophen can cause allergic acute TIN.

Immunological characterization of latent tuberculosis infection in a low endemic country

The diagnosis of a latent tuberculosis infection (LTBI) is based on detection of immunity against Mycobacterium tuberculosis (Mtb). The tuberculin skin test (TST), the Quantiferon (QFT) and a prolonged lymphocyte stimulation test using either ESAT-6/CFP-10 (LST-EC) or PPD (LST-PPD) were evaluated in a cohort of 495 individuals, suspected to have LTBI, in a low endemic country. While the TST and LST-PPD were both positive in the majority (75%) of individuals, only one third responded in the LST-EC and in the QFT. The choice for LTBI treatment was significantly associated with ESAT6/CFP10 recognition, however the LST-EC detected considerably more individuals (21%) with immunity against Mtb, who might also be at risk for development of active TB, although none of them did during follow up. Follow-up for 2 years showed 7% conversions and 32% reversions for the QFT. The LST-EC showed higher conversion rates (∼45%), although the percentage of individuals positive in the LST-EC did not change significantly within the follow-up period. LTBI treatment did not alter immune recognition of Mtb antigens. In conclusion, the sensitivity of tests for detection of cellular immunity to Mtb specific antigens depends on test methodology and may vary considerably over time in a low endemic region.

5-Aminosalicylate intolerance causing exacerbation in pediatric ulcerative colitis.

5-Aminosalicylate (5-ASA) is widely used as the first-line drug for ulcerative colitis (UC). 5-ASA is mostly a safe and effective drug, but it can bring about exacerbation due to 5-ASA intolerance. 5-ASA intolerance can be confusing and it can mislead physicians into considering unnecessary treatment escalation, including corticosteroid (CS), biologics, or even surgery. In spite of the clinical importance of 5-ASA intolerance, there have been few studies on its incidence, clinical features, and diagnosis. In order to evaluate the incidence, characteristic symptoms, disease course, and laboratory data of children with 5-ASA intolerance, we retrospectively reviewed the medical records of 80 children with UC. Eleven of 80 children (13.8%) with UC were diagnosed with 5-ASA intolerance. The median time between the initiation of 5-ASA and the onset of 5-ASA intolerance was 10days (range, 4-20days) in patients not receiving CS. Drug-induced lymphocyte stimulation test (DLST) was performed in 10 patients, and was positive in eight. C-reactive protein (CRP) increased significantly when exacerbation of colitis symptoms occurred. The incidence of 5-ASA intolerance was relatively high. Besides the challenge test, elevation of CRP and positive DLST appeared to support the diagnosis of 5-ASA intolerance.

Novel interferon-γ enzyme-linked immunoSpot assay using activated cells for identifying hypersensitivity-inducing drug culprits

BackgroundThe drug-induced lymphocyte stimulation test (DLST), also referred to as lymphocyte transformation test (LTT), is used to identify the culprit drug in cases of cutaneous adverse drug reactions (cADR). Although DLST is a widely used in vitro test, its sensitivity and specificity are unsatisfactory. Recent reports suggest that the detection of drug-induced interferon (IFN)-γ production using enzyme-linked immunoSpot (ELISpot) assay (conventional IFN-γ ELISpot) is useful for identifying culprit drugs in cADR cases. ObjectiveThe aim of this study was to establish a novel method for identifying culprit drugs in patients with cADR by efficiently detecting drug-specific IFN-γ production using activated cells. MethodsSixteen patients with cADR, including drug-induced hypersensitivity syndrome, erythema multiforme-like eruption, maculopapular exanthema, Stevens-Johnson syndrome, and toxic epidermal necrolysis, caused by clinically convincing culprit drugs were enrolled in this study. In some cases, the blood samples were obtained at two or three different time points. Peripheral blood mononuclear cells (PBMCs) from total 20 samples were analyzed using both the DLST and drug-induced conventional IFN-γ ELISpot. In addition, drug-induced IFN-γ ELISpot was performed using PBMCs, which were stimulated with anti-cluster of differentiation (CD)-3/CD28 antibody-coated microbeads and interleukin (IL)-2 for 7 days before exposure to the culprit drugs (modified IFN-γ ELISpot). ResultsAmong the culprit drugs tested in each patient, the modified IFN-γ ELISpot was positive in 17 samples (13 patients) while DLST and conventional IFN-γ ELISpot were positive in eight and four samples (six and three patients), respectively. ConclusionThe modified IFN-γ ELISpot using activated PBMCs was more sensitive than the conventional IFN-γ ELISpot was for detecting drug-induced IFN-γ production, which could be a useful in vitro tool for identifying culprit drugs in cADR cases.

Experimental infection with Salmonella enterica subsp. diarizonae serotype 61:k:1,5,(7) in sheep: Study of cell mediated immune response

The aim of the present study was to investigate the clinical, pathological and immunological changes in lambs infected experimentally with Salmonella enterica subsp. diarizonae serotype 61:k:1,5,(7) (SED) that causes chronic proliferative rhinitis (CPR) in sheep. Eleven four-month-old lambs were infected with 3.2×108UFC of SED by nasal route and four lambs with the same age were kept as placebo group. Clinical explorations were carried out monthly and vaginal, nasal swaps and fecal samples were taken for microbiological study during one year. Furthermore, a postmortem investigation was performed. Ventral turbinates of both nostrils and submandibular, parotid and retropharyngeal lymph nodes were sampled for histopathological, immunohistochemical and microbiological studies. Cell mediated immune response was assessed by lymphocyte stimulation test (Con A, PWM, PHA and LPS were used as mitogens), DTH test and by measuring IFN-γ concentration in serum and cultured supernatants.SED was isolated from nasal swabs and fecal samples of all infected lambs virtually throughout the study. No isolations were obtained from control group. At necropsy, no lesions associated with SED were recorded; however, the microorganism was isolated in regional lymph nodes in all infected animals. Moreover, infected lambs showed lower stimulation indices to Con A, PHA and PWM and higher for both LPS (P<0.001). Besides, DTH response was lower in infected lambs than in the control group, but statistical significant differences were not found (P>0.05). Serum levels of IFN-γ during the experiment were higher in infected sheep than in the control group, suggesting an active infection. Furthermore, mitogens stimulated the production of IFN-γ in lymphocyte cultures from infected sheep, and the highest level were obtained with Con A.These results demonstrate that after experimental infection of healthy lambs with S. enterica subsp. diarizonae serotype 61:k:1,5,(7) infection was maintained for at least one year in the upper respiratory tract and that digestive route should be taken into account as a possible way of SED elimination. Moreover, SED infection caused a weak response of the T cells; nevertheless, more studies are needed to understand the mechanisms involved in the immunitary response and pathology of CPR in sheep associated with Salmonella enterica subsp. diarizonae infection.

Induction of Remission and Maintenance Therapy with Tacrolimus in Refractory Ulcerative Colitis with Adverse Events from 5-ASA and Thiopurine

A 58-year-old Japanese woman was admitted to the hospital with fever, diarrhea, melena and lower abdominal pain. Colonoscopy showed moderate left-sided Ulcerative Colitis (UC). Administration of 5-aminosalicylic acid (5- ASA) (3600 mg) was started, but skin eruptions and hepatic dysfunction occurred. The drug lymphocyte stimulation test (DLST) was positive for 5-ASA, so this medication was discontinued. Prednisolone (PSL, initially 10 mg/day) was started, and remission was achieved. However, relapse occurred 5 months later after the discontinuation of PSL, and hospitalization was required again. Remission was achieved again by PSL, and azathioprine (AZA) (initially 25 mg/day) was started as maintenance therapy. Hepatic dysfunction was detected at 1 month after starting AZA and was judged to represent drug-induced hepatitis. Following the discontinuation of AZA, PSL (2 mg/day) monotherapy was continued. At 11 months after discharge from hospital, the patient developed diarrhea, melena, and abdominal pain, requiring hospitalization for the third time. Colonoscopy showed extensive ulceration and spontaneous bleeding, indicating a relapse of severe UC. Administration of tacrolimus (TAC) led to remission and the patient was discharged from hospital. Because she experienced adverse effects with 5-ASA and AZA, TAC alone was continued as maintenance therapy for ≥ 2 years after the discontinuation of PSL. Relapse did not occur during this period and repeat colonoscopy showed mucosal healing. During the TAC maintenance therapy, slight deterioration of renal function was observed, but there were no other adverse events. In conclusion, we experienced a rare patient who responded to TAC maintenance monotherapy for severe UC.

ロピバカインを用いた腹横筋膜面ブロック併用全身麻酔後に生じた薬剤性肝障害の1例

薬剤性肝障害の既往のある45歳女性に対し，腹式単純子宮全摘術が予定された．ロピバカインを用いた腹横筋膜面ブロック併用全身麻酔を施行した術後当日から，肝機能異常と凝固障害を認めた．循環不全やウイルスによる肝障害は否定的で，術中使用薬剤対象のリンパ球刺激試験（drug-induced lymphocyte stimulation test：DLST）でロピバカインが陽性と判定され，薬剤性肝障害が疑われた．ロピバカインでの肝障害の報告は少ないが，アレルギー性や代謝性といった特異体質性では薬剤性肝障害を発症しうる．本症例では，DLSTの結果から今後はロピバカインの使用は避けるべきであると考えられた．

Liver injury after aluminum potassium sulfate and tannic acid treatment of hemorrhoids.

We are reporting a rare case of acute liver injury that developed after an internal hemorrhoid treatment with the aluminum potassium sulfate and tannic acid (ALTA) regimen. A 41-year-old man developed a fever and liver injury after undergoing internal hemorrhoid treatment with a submucosal injection of ALTA with lidocaine. The acute liver injury was classified clinically as hepatocellular and pathologically as cholestastic. We could not classify the mechanism of injury. High eosinophil and immunoglobulin E levels characterized the injury, and a drug lymphocyte stimulation test was negative on postoperative day 25. Fluid replacement for two weeks after hospitalization improved the liver injury. ALTA therapy involves injecting chemicals into the submucosa, from the rectum to the anus, and this is the first description of a case that developed a severe liver disorder after this treatment; hence, an analysis of future cases as they accumulate is desirable.

薬剤性腎障害

薬剤性腎障害は薬剤の投与により新たに発症した腎障害，あるいは既存の腎障害の悪化を認める場合と定義される。腎は血流量が多いこと，薬物の濃縮が行われること，尿細管で分泌が行われその過程で尿細管細胞内の薬物濃度が上昇することなどにより，薬剤性障害が起こりやすい臓器である。薬剤性腎障害の診断は一般的な血液，尿検査，画像検査などと共に薬剤反応性リンパ球刺激試験(drug induced lymphocyte stimulation test: DLST)，尿中好酸球も含めて診断するが，薬剤性障害の診断基準に必ずしも当てはまらず，確定診断は困難である。予防や治療に特異的なものはなく，尿の濃縮を防ぐための輸液負荷，薬剤の減量などしかない。

Interstitial pneumonia in a patient treated with TAS-102 for metastatic colorectal cancer: a case report

BackgroundTAS-102, a new treatment option for patients with metastatic colorectal cancer that is refractory or intolerant to standard therapies, has been improving survival with acceptable tolerability and adverse events. Adverse hematological events associated with TAS-102 treatment were extensively profiled in the RECOURSE trial, but pulmonary toxicities associated with TAS-102 therapy are distinctly uncommon. In a recent early post-marketing phase vigilance on TAS-102 in Japan, seven cases of pulmonary disease were reported, but patient follow-up in this study was incomplete. Here, we present the first case of interstitial lung disease occurring in association with TAS-102 treatment.Case presentationA 57-year-old Japanese man who had previously received two standard treatments was admitted in 2014, at which time we administered TAS-102 (110 mg/day) as a third-line chemotherapy. He was safely treated with TAS-102 for the first planned cycle; however, approximately 4 days after receiving the second cycle of TAS-102, he complained of high fever and subsequent dyspnea with severe hypoxemia and went to the emergency room. A chest X-ray revealed diffuse coarse reticular shadows with ground-glass opacity on both lungs. Furthermore, a chest computed tomography scan showed thickening of the bronchovascular bundles with extensive ground-glass opacification and pleural effusions in both lung fields.In addition, a peripheral blood lymphocyte stimulation test with TAS-102 showed higher values compared with control samples. Consequently, we suspected drug-induced interstitial pneumonia, and discontinued treatment. Our patient was given an initial administration of high-dose methylprednisolone (1000 mg/day) for 3 days and oxygen. Our patient was discharged with oral prednisolone (20 mg/day) and improved symptomatically and radiologically.ConclusionsThese findings suggest that interstitial pneumonia is a rare complication of TAS-102 chemotherapy, but the possibility of interstitial pneumonia should always be considered when a patient presents with a respiratory disorder while undergoing TAS-102 systemic chemotherapy.Prompt discontinuation of TAS-102 and treatment with high-dosage corticosteroids is needed to avoid exacerbating respiratory symptoms.

気管支肺胞洗浄液での薬剤リンパ球刺激試験陽性を認めたTS-1^[○!R]による薬剤性肺障害の1例

気管支肺胞洗浄液での薬剤リンパ球刺激試験陽性を認めたTS-1^[○!R]による薬剤性肺障害の1例

398 Novel lymphocyte stimulation tests using ex-vivo expanded skin-infiltrating T cells from severe drug eruptions

398 Novel lymphocyte stimulation tests using ex-vivo expanded skin-infiltrating T cells from severe drug eruptions

How safe is the use of chlorpyrifos: Revelations through its effect on layer birds.

Aim:The present study was aimed to investigate the immunological competence of chlorpyrifos (CPF) insecticide after oral administration in layer chickens.Materials and Methods:A total of 20 White Leghorn birds were given CPF in drinking water at 0.3 ppm/bird/day (no observable effect level dose) for a period of 3-month. Immune competence status of layer birds and chicks hatched from CPF-treated birds were estimated at 15 days interval in layer birds and monthly interval in chicks using immunological and biochemical parameters.Results:There was a significant decrease in values of total leukocytes count, absolute lymphocyte count, absolute heterophil count, total serum protein, serum albumin, serum globulin, and serum gamma globulin in the birds treated with CPF as compared to control. Similarly, immune competence tests such as lymphocyte stimulation test, oxidative burst assay, and enzyme-linked immunosorbent assay tests indicated lower immunity in birds treated with CPF as compared to control. Subsequently, chicks produced from CPF-treated birds were also examined for immune competence, but no significant difference was observed between chicks of both the groups.Conclusion:The exposure to CPF produced hemo-biochemical and other changes that could be correlated with changes in the immunological profile of layer chickens suggesting total stoppage of using CPF in poultry sheds.

Effect of endosulfan on immunological competence of layer birds.

Aim:The present study was aimed to investigate the immunological competence of endosulfan insecticide after limited oral administration in White Leghorn layer chickens.Materials and Methods:A total of 20 White Leghorn birds were given endosulfan in drinking water at 30 ppm/bird/day (no observable effect level dose) for a period of 3-months. Immune competence status of layer birds and chicks hatched from endosulfan offered birds were estimated at 15-day interval in layer birds and at monthly interval in chicks using immunological, biochemical parameters, and teratological estimates.Results:There was a significant decrease in levels of total leukocytes count, absolute lymphocyte count, absolute heterophil count, total serum protein, serum albumin, serum globulin, and serum gamma globulin in the birds fed with endosulfan as compared to control. Similarly, immune competence tests such as lymphocyte stimulation test, oxidative burst assay, and enzyme-linked immunosorbent assay tests indicated lower immunity in birds treated with endosulfan as compared to control. Subsequently, chicks produced from endosulfan-treated birds were also examined for immune competence, but no significant difference was observed between chicks of both the groups.Conclusion:The exposure to endosulfan in limited oral dosage was able to exhibit hemo-biochemical and other changes that could be correlated with changes in the immunological profile of layer chickens suggesting cautious usage of endosulfan insecticide in poultry sheds.

Immune response in adverse reactions to metal debris following metal-on-metal total hip arthroplasty

BackgroundThe purpose of the present study was to determine whether T cell-mediated type IV hypersensitivity reactions could be a major cause of adverse reaction to metal debris (ARMD) after metal-on-metal total hip arthroplasty (THA).MethodsThirteen patients (1 man and 12 women; mean age 68 years, age range 60 to 83 years) with ARMD underwent revision surgery following metal-on-metal THA (15 hips). Lymphocyte stimulation testing was conducted. Periprosthetic tissue specimens underwent immunohistochemical studies.ResultsLymphocyte stimulation testing showed that five patients were nickel-sensitive, and one patient was also cobalt-sensitive. Immunohistochemical studies showed that T cells were dominant in five hips, and B cells were dominant in 10 hips. In four of the five patients with a positive lymphocyte stimulation test, the dominant lymphocytes were T cells, suggesting type IV hypersensitivity. The major cause of ARMD was not type IV hypersensitivity in the remaining nine patients.ConclusionMetal hypersensitivity does not appear to be the dominant biological reaction involved in the occurrence of ARMD.Electronic supplementary materialThe online version of this article (doi:10.1186/s12891-016-1069-9) contains supplementary material, which is available to authorized users.