Specific Immunotherapy Treatment Research Articles

Occupational asthma and rhinitis due to wheat flour: sublingual specific immunotherapy treatment.

Background:There are several potential sensitizers in the bakery environment and wheat flour appears to be the dominant sensitizer in most bakeries. Apart from traditional drug therapy or a change in profession, there are no effective therapies for workers who develop serious respiratory symptoms in the workplace.Objectives:To describe clinical and laboratory findings in workers with asthma and/or rhinitis induced by wheat flour who underwent sublingual specific immunotherapy (SLIT).Methods:Since drug therapy and prevention strategies were not effective, five bakers were elected to undergo SLIT. A three-year study was led by administering a sublingual wheat flour extract. Questionnaires, allergy and respiratory tests were performed before and after SLIT.Results:After SLIT an improvement in symptoms is observed in every patient: Asthma Control Test and a quality-of-life questionnaire show higher scores and as a result, workers have reduced the use of drug therapy. We observed significantly reduced exhaled nitric oxide (FeNO) and eosinophil cationic protein (ECP) levels after SLIT, hypothesizing that these parameters may be used to monitor the effectiveness of immunotherapy. The improvement of FEV1 (forced expiratory volume in 1second) and responsiveness to bronchoprovocative tests with methacholine denotes a possible role of SLIT in treating patients with low-respiratory tract involvement, even though more data are needed.Discussions:This is the first report in the literature on the use of SLIT for baker’s asthma and rhinitis. SLIT for occupational wheat flour allergy should be possible and efficient, saving vocational training, professionalism, and avoiding job loss.

Abstract 4049: Assessment of immune biomarkers by digital pathological analysis across a large colorectal cancer patient cohort predicts patient outcome and may provide a clinically relevant therapeutic index for immunotherapeutic treatment stratification

Abstract Introduction: Colorectal cancer (CRC) is a heterogeneous malignancy, treatment for which has principally been cytotoxic chemotherapy. Herein we describe the assessment of immune-biomarkers in a large patient cohort, demonstrating the therapeutic potential of immunotherapy in CRC. Methods: By immunohistochemistry, employing REMARK criteria, using well-validated antibodies, 4 biomarkers (CD3, CD4, CD8, FOXP3) in over 1000 cases, represented across 33 TMAs, were assessed for their protein expression. Digital pathological assessment of immune biomarker densities were quantified via the application of QuPath, an open source digital pathology platform, developed at Queens University, Belfast (https://qupath.github.io.). Biomarker densities were dichotomised by R.O.C analysis. Kaplan-Meier curves (Log-rank p values) were used to assess the impact of immune cell densities on overall survival (OS) and progression-free survival (PFS). Hazard ratios were assessed using the Cox proportional hazards regression model for univariate analysis. Spearman's rank was used to assess correlations of the biomarkers. Results: Expression of two biomarkers were found to vary by stage and three significantly affected OS. Across the patient cohort CD3, CD4 and CD8 were individually shown to be positively prognostic for OS alone (log-rank p = 0.0007, p = 8.65e-06, p = 0.006, respectively). CD4 demonstrated a more significant impact on OS in later stage disease (Log-rank stage II p = 0.0017, stage III p = 0.0002), while conversely CD8 only showed significance in earlier stage disease (Log-rank stage II p = 0.009, stage III p = 0.546). Although significantly associated with OS and within all stages, CD3 showed no differential expression in a particular stage. Correlative analysis demonstrated a statistically significant association between biomarkers. Upon univariate analysis, all biomarkers remained significantly prognostic for OS. All analyses were performed using R. Conclusions: These data demonstrate the prognostic significance of adaptive immune markers in a large CRC cohort on OS. Moreover, stage specific expression maybe clinically relevant and assessment within stages II and III may provide important stratifications for specific immunotherapy treatment regimes. We additionally add weight to the beneficial utility of digital pathology and its potential for integration into routine tissue biomarker assessment. Additional analysis of immune checkpoint and adaptive immune biomarkers in CRC is warranted. Citation Format: Matthew P. Humphries, Stephanie Craig, Victoria Bingham, Maurice Loughrey, Helen Coleman, Liam Murray, Maughan Tim, Stephen McQuaid, Jacqueline James, Manuel Salto-Tellez. Assessment of immune biomarkers by digital pathological analysis across a large colorectal cancer patient cohort predicts patient outcome and may provide a clinically relevant therapeutic index for immunotherapeutic treatment stratification [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4049.

Safety of immunotherapy with glutaraldehyde modified allergen extracts in children and adults

BackgroundGlutaraldehyde-modified natural allergen extracts show significant reduction in the IgE-binding capacity and proteolytic activity. This allows the administration of higher doses in a shorter period of time, and to mix different allergen extracts. ObjectiveEvaluate the safety of different concentrations and mixtures of glutaraldehyde-modified allergen extracts in a large group of paediatric and adult patients undergoing specific immunotherapy treatment. Materials and methods1855 patients (1156 adults and 699 children), suffering from rhinoconjunctivitis and/or asthma, participated in an observational multicentre cohort study, to evaluate the safety of immunotherapy using vaccines containing modified allergen extracts. Patients were monosensitised, or polysensitised, and received a therapeutic vaccine containing polymerised allergen extracts adsorbed onto aluminium hydroxide. Safety was assessed by recording all side reactions related to immunotherapy. ResultsThe clinically relevant local reactions totalled 120, (90 immediate and 30 delayed) (1.02% of injections). Of them, 31 (0.26% of injections) occurred in children (26 immediate and 5 delayed) and 89 (0.76% of injections) in adults (64 immediate and 25 delayed). There were 38 systemic reactions. Eleven reactions were immediate (9 of grade 1 and 2 of grade 2) and 27 delayed (22 of grade 1 and 5 of grade 2). There were seven grade 2 systemic reactions (0.06% of the injections). No differences (P>0.05) in the number of reactions were observed between adults and children and between treatments were found in systemic reactions. All systemic reactions were mild and resolved spontaneously without the need of medication. ConclusionSpecific immunotherapy using natural modified allergen vaccines is safe to treat allergic patients, even at higher doses and in mixtures of unrelated allergen extracts. The percentage of adverse reactions detected is lower than those reported in the literature with native unmodified allergen extracts.

Measurement of serum antibodies against NY-ESO-1 by ELISA: A guide for the treatment of specific immunotherapy for patients with advanced colorectal cancer.

NY-ESO-1 has been identified as one of the most immunogenic antigens; thus, is a highly attractive target for cancer immunotherapy. The present study analyzed the expression of serum antibodies (Abs) against NY-ESO-1 in patients with advanced colorectal cancer (CRC), with the aim of guiding the treatment of NY-ESO-1-based specific-immunotherapy for these patients. Furthermore, the present study was the first to evaluate the kinetic expression of anti-NY-ESO-1 Abs and investigate the possible influencing factors. A total of 239 serum samples from 155 pathologically confirmed patients with advanced CRC (stages III and IV) were collected. The presence of spontaneous Abs against NY-ESO-1 was analyzed using an enzyme-linked immunosorbent assay (ELISA). The results demonstrated that 24.5% (38/155) of the investigated patients were positive for NY-ESO-1-specific Abs. No statistically significant correlations were identified between the expression of anti-NY-ESO-1 Abs and clinicopathological parameters, including age and gender, location, grading, local infiltration, lymph node status, metastatic status and K-ras mutation status (P>0.05). In 59 patients, the kinetic expression of anti-NY-ESO-1 Abs was analyzed, of which 14 patients were initially positive and 45 patients were initially negative. Notably, 16/59 (27.1%) patients changed their expression status during the study period, and the initially positive patients were more likely to change compared with the initially negative patients (85.7 vs. 8.8%; P<0.001). Therefore, monitoring serum Abs against NY-ESO-1 by ELISA is an easy and feasible method. The high expression rate of NY-ESO-1-specific Abs in CRC patients indicates that measuring the levels of serum Abs against NY-ESO-1 may guide the treatment of NY-ESO-1-based specific immunotherapy for patients with advanced CRC.

Omalizumab for the Treatment of Inadequately Controlled Allergic Rhinitis: A Systematic Review and Meta-Analysis of Randomized Clinical Trials

Patients with moderate-to-severe allergic rhinitis who are inadequately controlled despite treatment according to current rhinitis management guidelines have a significant unmet medical need. Such patients have a negative impact on daily functioning and are at risk of developing serious comorbidities, such as asthma and chronic rhinosinusitis. To assess the efficacy and safety of omalizumab in poorly controlled allergic rhinitis under a meta-analysis framework. MEDLINE and the Cochrane Central Register of Controlled Trials were searched through September 2013. Studies on the efficacy of omalizumab in allergic rhinitis that assessed clinical outcomes were selected. Descriptive and quantitative information was extracted; mean differences and relative risk estimates were synthesized under a fixed or random effects model. Heterogeneity was assessed by using the Q statistic and the I(2) metric. Subgroup analyses were performed for the presence of specific immunotherapy treatment. Of the 352 citations retrieved, 11 studies of 2870 patients were finally included. A statistically significant reduction in the daily nasal symptom severity score (standardized mean difference-0.67 [95% CI,-1.3 to-0.31]; P < .0001; I(2), 92%) and a statistically significant reduction in daily nasal rescue medication score (-0.22 [95% CI,-0.39 to-0.05; P= .01; I(2), 58%) were observed. There was not a statistically significant difference in the occurrence of any adverse event (relative risk 1.06 [95% CI, 0.94-1.19; I(2), 55%). Omalizumab is statistically significantly associated with symptom relief, decreased rescue medication use, and improvement of quality of life in patients with inadequately controlled allergic rhinosinusitis.

Impact of allergic rhinitis and specific subcutaneous immunotherapy on peripheral blood basophils of patients sensitized to Dermatophagoides pteronyssinus

BackgroundBasophils are important effectors cells in allergic rhinitis (AR) since they are involved in immunoglobulin (Ig) E – mediated inflammation and in the release of pro-inflammatory mediators. Specific subcutaneous immunotherapy (SCIT) provides clear immunologic modulation in some immune cells, however its systemic effects on basophils are not well known.MethodsPeripheral blood (PB) samples from 43 patients with allergic rhinitis mono-sensitized to Dermatophagoides pteronyssinus (Dpt) [33 of them under SCIT with allergoid Dpt extract, in maintenance dose (SCIT), with evaluation just before SCIT injection (SCIT-T0) and 4 hours later (SCIT-T4) and the other 10 Dpt allergic patients never having, in the past, undergone specific immunotherapy treatment (NSIT)], and 15 healthy age- and gender-matched controls (HG), were analyzed. For each sample, the total (t-IgE) and specific IgE (s-IgE) was performed, as well as, the relative frequency and absolute number of PB basophils and receptor-bound IgE and IgG expression were evaluated by flow cytometry and the Histamine N-methyltransferase (HNMT) and tryptase α/β1 (TPSAB1) gene expression was assessed by real-time PCR.ResultsHigher levels of receptor-bound IgE were observed in SCIT patients, which are correlated with the levels of serum t-IgE and s-IgE, whereas no significant differences were observed for receptor-bound IgG. Regarding HNMT mRNA expression, significantly lower expression levels were detected in AR patients compared to HG, independently of type of therapy. Moreover a negative correlation was found between HNMT gene expression and time under SCIT. Conversely, tryptase gene expression was significantly up-regulated in NSIT when compared to HG; however in SCIT patients, tryptase gene expression was significantly decreased than in NSIT patients. No differences were found for any parameter between SCIT-T0 and SCIT-T4 with exception of a transient increased expression of tryptase in SCIT-T4.ConclusionPB basophils from patients with AR show altered functional features, which seems to be influenced by SCIT, suggesting that these cells could be useful to clarify the SCIT triggered mechanisms at a systemic level.

T cell responses to known allergen proteins are differently polarized and account for a variable fraction of total response to allergen extracts.

A panel of 133 allergens derived from 28 different sources, including fungi, trees, grasses, weeds, and indoor allergens, was surveyed utilizing prediction of HLA class II-binding peptides and ELISPOT assays with PBMC from allergic donors, resulting in the identification of 257 T cell epitopes. More than 90% of the epitopes were novel, and for 14 allergen sources were the first ever identified to our knowledge. The epitopes identified in the different allergen sources summed up to a variable fraction of the total extract response. In cases of allergens in which the identified T cell epitopes accounted for a minor fraction of the extract response, fewer known protein sequences were available, suggesting that for low epitope coverage allergen sources, additional allergen proteins remain to be identified. IL-5 and IFN-γ responses were measured as prototype Th2 and Th1 responses, respectively. Whereas in some cases (e.g., orchard grass, Alternaria, cypress, and Russian thistle) IL-5 production greatly exceeded IFN-γ, in others (e.g., Aspergillus, Penicillum, and alder) the production of IFN-γ exceeded IL-5. Thus, different allergen sources are associated with variable polarization of the responding T cells. The present study represents the most comprehensive survey to date of human allergen-derived T cell epitopes. These epitopes might be used to characterize T cell phenotype/T cell plasticity as a function of seasonality, or as a result of specific immunotherapy treatment or varying disease severity (asthma or rhinitis).

4 The Risk of Severe Treatment Related Adverse Events is Significantly Lower in Children Compared to Adults When Treated With Standardized Timothy Grass Allergy Immunotherapy Tablets

BackgroundAllergy immunotherapy tablets (AIT) provides a safer and more convenient alternative to subcutaneous specific immunotherapy treatment. However, severe adverse events occur infrequently. These events are rare and therefore pooled safety data from 1 phase II/III and 6 phase III clinical trials (5 in adults, 2 in children (5–17 years)) with grass AIT (Grazax, Phleum pratense 75,000 SQ-T/2800 BAU, ALK, Denmark) were analysed.MethodAll trials were randomised, double-blind, placebo-controlled multi-centre trials. Subjects suffered from grass pollen induced allergic rhinoconjunctivitis with or without asthma, had positive skin prick test and specific IgE to Phleum pratense. Subjects received once-daily sublingual treatment with grass AIT or placebo for approximately 24 weeks. The 5 adult trials comprised 2095 treated subjects (AIT = 1060, placebo = 1035) and the 2 children trials comprised 597 treated subjects (AIT = 301, placebo = 296). Adverse events were assessed by the investigator as treatment-related (possible or probable) or unlikely related and for seriousness. Application site-related events were defined as adverse events in relation to the oral cavity.ResultsIn the adult trials, 71% of AIT-treated subjects reported treatment-related adverse events compared to 24% for placebo. In the children trials the corresponding numbers were 63% for AIT and 27% for placebo. Of the AIT-treated subjects 2 children (0.7%) and 32 adults (3.0%) experienced severe treatment-related events. The odds for severe events was 4.7 times lower in children compared to adults (odds-ratio with 95% CI, 0.22 [0.025-0.85], P = 0.019). Both AIT-treated children (0.7%) and 18 (1.7%) of the AIT-treated adults experienced severe treatment-related events that were application site-related. The odds for having severe related application site adverse events was 2.6 times lower in children compared to adults (odds-ratio with 95% CI, 0.39 [0.04-1.63], P = 0.27, not statistically significant). No serious treatment-related adverse events were reported.ConclusionThis pooled analysis of over 2000 subjects in 7 clinical trials shows that the risk of experiencing severe treatment-related adverse events was significantly lower in children compared to adults when treated with of Timothy grass allergy immunotherapy tablets. This analysis provides evidence that Timothy grass AIT is an important and safe immunotherapy treatment option in children with grass pollen induced rhinoconjunctivitis.

Abstract 897: Thymidine kinase 1, a novel biomarker specific to the plasma membrane of cancerous cell lines

Abstract Thymidine Kinase 1 (TK1), an important nucleotide salvage pathway enzyme, has been shown to be upregulated in most malignant cancers. Recent studies have shown that TK1 not only has diagnostic and prognostic potential, but could also be used as a molecular target for therapeutic treatments. Previous research from our lab has shown the expression of TK1 on the plasma membrane of lymphoma. This study provides further insight on the role of TK1 as a diagnostic target for several different tumor cell lines. This study analyzed the localization of TK1 in ten different cancer cell lines using flow cytometry (FCM). The lines studied were A375, a malignant melanoma, CAL27, a squamous cell carcinoma derived from tongue, MDA-MB-231, a breast adenocarcinoma, MDA-MB-435S, previously described as a ductal carcinoma, HT29, a colorectal adenocarcinoma, SW620, a colorectal adenocarcinoma, A549, a lung carcinoma, NCI-H460, a large cell lung carcinoma, DU 145, a prostrate carcinoma, and PC3, a prostate adenocarcinoma. FCM was carried out using a polyclonal antibody against human TK1 as the primary antibody and a FITC conjugated secondary antibody. A sample stained with only secondary antibody was used as a negative control for the experiment. Five replicates were analyzed for each cell line. TK1 was found to bind between 48% and 96%, depending on the cell line. Non-cancerous tonsil tissue and normal lymphocytes were used as controls, where TK1 surface binding was less than 20%. These data indicate the feasibility of TK1 as a possible molecular target for immunotherapy in many types of cancer. Further research is ongoing to determine specific immunotherapy treatment options, and to verify the expression of TK1 in other cancer cell lines. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 897. doi:10.1158/1538-7445.AM2011-897

Molecular determinants of T cell epitope recognition to the common Timothy grass allergen.

We investigated the molecular determinants of allergen-derived T cell epitopes in humans utilizing the Phleum pratense (Timothy grass) allergens (Phl p). PBMCs from allergic individuals were tested in ELISPOT assays with overlapping peptides spanning known Phl p allergens. A total of 43 distinct antigenic regions were recognized, illustrating the large breadth of grass-specific T cell epitopes. Th2 cytokines (as represented by IL-5) were predominant, whereas IFN-gamma, IL-10, and IL-17 were detected less frequently. Responses from specific immunotherapy treatment individuals were weaker and less consistent, yet similar in epitope specificity and cytokine pattern to allergic donors, whereas nonallergic individuals were essentially nonreactive. Despite the large breadth of recognition, nine dominant antigenic regions were defined, each recognized by multiple donors, accounting for 51% of the total response. Multiple HLA molecules and loci restricted the dominant regions, and the immunodominant epitopes could be predicted using bioinformatic algorithms specific for 23 common HLA-DR, DP, and DQ molecules. Immunodominance was also apparent at the Phl p Ag level. It was found that 52, 19, and 14% of the total response was directed to Phl p 5, 1, and 3, respectively. Interestingly, little or no correlation between Phl p-specific IgE levels and T cell responses was found. Thus, certain intrinsic features of the allergen protein might influence immunogenicity at the level of T cell reactivity. Consistent with this notion, different Phl p Ags were associated with distinct patterns of IL-5, IFN-gamma, IL-10, and IL-17 production.

Russian thistle pollinosis: form allergen characterization to specific immunotherapy treatment

Chenopodiacea/Amaranthacea pollen is an important cause of respiratory allergy around the world. The problem could be aggravated because in desert or desert-like countries, these weeds are used in greening programmes or as ornamental plants. As climate tendencies should maintain in future decades, relevance of Chenopodiacea/Amaranthacea pollen allergy will go up. Relationship between airborne pollen grains and symptoms may not follow conventional patterns, and a wide lag between pollen counts and symptoms has been observed. Allergens from Chenopodium album y Salsola kali are probably the best known. A mayor band protein of 40-43 kDa has been associated with the main Salsola kali allergen, named sal k 1 and now recognised as pectin methyltranspherase or glucose 3 phosphate dehidrogenase. Systemic immunotherapy is effective and safe, improving symptoms, medication intake and Quality of Life and also reduces cutaneous allergen response.

Comparison of the Long-Term Efficacy of Subcutaneous and Sublingual Immunotherapies in Perennial Rhinitis

Background: Both sublingual and subcutaneous immunotherapies have a documented clinical efficacy, but only a few comparative studies have been performed. Objectives: To evaluate and compare the long-term efficacies of subcutaneous and sublingual immunotherapy. Methods: One hundred and ninety-three patients with house dust mite allergies, out of an original total of 230, were treated with subcutaneous and sublingual house dust mite-specific immunotherapies for 3 years and also observed for 3 years after discontinuation of the treatment. The patients were randomized into 2 groups: the sublingual group (97 patients) and the subcutaneous group (96 patients). The therapy’s success was evaluated using the symptom score, skin prick test results, and the nasal allergen challenge score. The patients were evaluated at the beginning of the study, at the end of years 1, 2, and 3, and also at the end of the 1st and 3rd years after discontinuation of the specific immunotherapy treatment. Results: Immunotherapy induced a significant improvement during the treatment and the follow-up period. We found a greater improvement in the subcutaneous group compared to the sublingual group when we looked at the comparative results of the total 6 years. Conclusion: We suggest subcutaneous immunotherapy for patients with perennial allergic rhinitis due to the better results that were obtained during our study period. Nevertheless, sublingual immunotherapy is now accepted by WHO as a valid alternative to the subcutaneous route and should be used in all patients who require immunotherapy and do not accept the subcutaneous route of allergen administration.

Utilización de inmunoterapia específica en la Zona Básica de Salud Campiña Sur (Moguer-Huelva)

To analyse the appropriateness of specific immunotherapy treatment (IT) to indications based on scientific evidence. Cross-sectional, descriptive study. Primary care in a health district of about 12,000 inhabitants. 166 patients for whom IT was prescribed in the period between January 1998 and June 1999 were included. The full evaluation covered 65 patients whose allergy report was available in their clinical records. IT prescription was considered adequate in patients with single sensitivity to respiratory allergy or anaphylaxis to insect poison, in whom well-standardised single-component "vaccines" were used. IT was only used adequately in 36.4% of cases (71.4% in asthma, 46.1% in naso-bronchial allergy, and 18.7% in rhinitis). The high prescription of accompanying medication should be highlighted: oral H1 anti-histamines (81.8%), inhaled steroids for asthma (42.4%) and nasal steroids for rhinitis (72.4%). Use of IT in our health district is poorly suited to the indications based on contrasted scientific evidence. Clinical practice guidelines must be agreed at all care levels so as to improve the quality of care for all allergy patients.

Antihistamine premedication in specific cluster immunotherapy: A double-blind, placebo-controlled study

BACKGROUND: Specific immunotherapy treatment in allergic diseases involves a risk of systemic side effects. A double-blind, placebo-controlled study was performed in 45 patients allergic to pollen to determine whether pretreatment with loratadine could reduce the number and severity of systemic reactions during the dose-increase phase of cluster immunotherapy. METHODS: The patients received cluster immunotherapy with a standardized birch (Betula verrucosa) or grass (Phleum pratense) pollen extract adsorbed to aluminum hydroxide. The immunotherapy schedule involved seven visits and 14 injections to reach a maintenance dose of 100,000 standardized quality units. Loratadine, 10 mg, or placebo tablets were administered 2 hours before the first injection at each visit. RESULTS: A total of 720 injections were given (309 injections in 21 patients receiving loratadine and 411 injections in 24 patients receiving placebo). The median numbers of injections to reach maintenance dose were 15 (range, 14 to 18) in the loratadine group and 16 (range, 14 to 23) in the placebo group (p = 0.037). The numbers of patients with systemic reactions were seven (33%) and 19 (79%) in the loratadine and placebo groups, respectively (p = 0.002). Twenty-five reductions caused by systemic reactions were observed in the placebo group in contrast to nine in the loratadine group (p = 0.047). No life-threatening systemic reactions were observed in either group. Systemic reactions were, however, more severe in the placebo group, mainly because of a significantly higher incidence of urticaria (10 vs 1, p = 0.022). CONCLUSION: Pretreatment with loratadine seems to reduce both the number and severity of systemic reactions in specific cluster immunotherapy. (J Allergy Clin Immunol 1996;97:1207-13.)