Strength Of Reactivity Research Articles

Warm reactive autoantibodies detected by solid phase technology: Patient characteristics, laboratory features, and clinical significance.

This study focused on the serology, clinical characteristics, and hemolytic potential of warm reactive autoantibodies detected by solid phase red cell adherence. Ninety-seven patients with warm autoantibodies were evaluated. Serologic characteristics included the strength of solid phase reactivity, the results of tube-based ancillary testing, direct antiglobulin test and eluate results, and an assessment for contemporaneous alloantibodies. Clinical characteristics of the patients included age, sex, and primary diagnosis. Each patient was also assessed for evidence of hemolysis. Most of the 97 study patients were female (63.9%), and the average age was 66 years. Hematologic disorders were the most common diagnosis. A majority (70.1%) of the warm autoantibodies had 3 to 4+ reaction strengths, and approximately 90% had negative testing with at least 1 test tube method. There was an even distribution of direct antiglobulin test reaction strengths, with 74% reactive with anti-immunoglobulin G only. Alloantibodies were identified in 20% of patients. Evidence of hemolysis was identified in only 13 patients (13.4%). Warm reactive autoantibodies are more likely to be hemolytic, have strongly reactive indirect and direct antiglobulin tests, remain reactive in tube-based ancillary testing methods, and are seen primarily in patients with hematologic disorders.

Evaluation of acid mine drainage (AMD) from tailings and their valorization by copper recovery

Recent investigations have found that the mining industry generates approximately 10 billion tons of tailings per year, tending towards doubling this amount by 2035. For this reason, a systematic method is needed to study acid mine drainage (AMD) potential, as well as determining the dissolution of potentially toxic elements (PTEs). Furthermore, the possible recovery of base or precious metals could transform the tailings into value-added materials. In the present investigation, the potential for the generation of acid mine drainage in a sample of mine tailings was evaluated, as well as an analysis of the dissolution of PTEs through a sequential extraction of appropriate chemical solutions with increasing strength of reactivity to identify base or precious metals that can be recovered through a metallurgical process consisting of leaching, solvent extraction and electrolysis, operating at moderate pressure and temperature. The results of this research showed that these tailings contain approximately 3.7% of pyrite occluded in silicates and therefore are not a high risk for generating acid drainage and PTEs dissolution. On the other hand, a process for obtaining metallic copper was proposed, which resulted in a leaching liquor with 1226 ppm using 0.21 M H2SO4 and a pulp density of 300 g/L. Solvent extraction in a single stage allowed the leach liquor to be concentrated at 9200 mg Cu/L, recovering the majority of the copper in metallic form by electrolysis.

Sarcopenia in the elderly versus microcirculation, inflammation status, and oxidative stress: A cross-sectional study

Age-related mechanisms of sarcopenia associated with vascular function have been recently suggested. This study compared and tested associations between muscle mass and strength, microcirculation, inflammatory biomarkers, and oxidative stress in older adults classified as sarcopenic and non-sarcopenic. Thirty-three physically inactive individuals (72±7 yrs) were assigned to age-matched sarcopenic (SG) and non-sarcopenic (NSG) groups. Between-group comparisons were performed for appendicular skeletal mass (ASM), handgrip and isokinetic strength, microvascular function and morphology, C-reactive protein, insulin-like growth factor-1, tumor necrosis factor-alpha, interleukin-6 (IL-6), soluble vascular cell adhesion molecule-1, soluble intercellular adhesion molecule-1, endothelin-1, and oxidized low-density lipoprotein. ASM and knee isokinetic strength were lower in SG than NSG (P < 0.05). No difference between groups was found for outcomes of microvascular function and morphology, but log-transformed IL-6 concentration was twice greater in SG vs. NSG (P = 0.02). Correlations between ASM index, handgrip and knee isokinetic strength vs. markers of microcirculatory function, capillary diameters, vascular reactivity, and endothelial injury were found only in SG. Decreased ASM index and strength have been associated with microcirculatory profile, indicating that microcirculation impairment may be involved somehow in Sarcopenia development. The inflammation status, particularly elevated IL-6, seems to play an important role in this process.

Analysis of the influence of coke quality on the production of pig iron in the conditions of JSC “Ural Steel”

The purpose of the study was to conformity of the quality of coke used in the blast-furnace shop of JSC “Ural Steel” meets the requirements recommended in the scientific literature, and to study its impact on the consumption of coke and the production of pig iron at the enterprise. For the study, the blast furnace № 1 with a useful volume of 1007 m3 and the blast furnace No. 4 with a useful volume of 2002 m3 were selected. The results of the analysis of data on the quality of coke over a three-year period: ash content, volatile matter and sulphur in the coke, the moisture content in the coke, the coke uniformity coefficient, indicators of the cold strength of coke (M25 and M10), indicators of coke hot strength (CSR) and reactivity of coke after high-temperature interaction with CO2 (CRI). Studies have revealed significant instability of the moisture content in the coke, which affects the change in its consumption, and heterogeneity of the granulometric composition of the coke, increasing the stability and uniformity of which would favorably affect the indicators of the blast furnace process. It was found that the values of the indicators of M10 coke attrition, hot strength of coke (CSR) and reactivity of coke (CRI) do not meet the recommendations of domestic and foreign scientists. Measures are proposed to improve the quality of coke, the implementation of which will increase the productivity of blast furnaces and reduce the consumption of coke per ton of pig iron.

Identification of RHD allelic variants discovered by atypical typing results on the NEO/Echo platforms

Abstract Some individuals are at risk of anti-D alloimmunization if they inherit D antigens that are qualitatively and/or quantitatively different than wild-type D. We hypothesized that patients who showed serologically inconsistent, weak, or historically discordant D typing results by microplate direct agglutination (MDA) on NEO or Echo (Immucor, Norcross, GA) might be at risk of carrying RHD allelic variants. The present study was designed to evaluate patients with RHD allelic variants if they presented with weakly reactive D typing results on the NEO or Echo. Patients were selected for RHD genotyping if their specimens showed weak reactivity with either series 4 or series 5 anti-D typing reagent, if the strength of reactivity was ≤1+ on the NEO or Echo, or if historical or current D typing results were discordant with current results. Patients selected for RHD genotyping were also tested by saline tube testing using the same anti-D series 4 and 5 reagents. Genotyping was performed by the Immucor genotyping laboratory in Warren, NJ. Of 80 patients whose samples met study inclusion, 52 (65.0%) were found to have RHD allelic variants. Sixteen patients (20.0%) expressed possible Ceppellini effect reactivity. Most importantly, 51.25 percent of the patients who presented with weakly reactive D typing results by MDA testing on the NEO (≤1+) or Echo (≤1+) had RHD allelic variants that were associated with the potential for anti-D alloimmunization. Laboratories that use MDA testing on the Neo or Echo for D typing should consider that female patients of childbearing age might be at risk of anti-D alloimmunization if they are classified as D+ based on weakly reactive D typing results.

Molecular T-Cell Repertoire Analysis as Source of Prognostic and Predictive Biomarkers for Checkpoint Blockade Immunotherapy.

The T cells are key players of the response to checkpoint blockade immunotherapy (CBI) and monitoring the strength and specificity of antitumor T-cell reactivity remains a crucial but elusive component of precision immunotherapy. The entire assembly of T-cell receptor (TCR) sequences accounts for antigen specificity and strength of the T-cell immune response. The TCR repertoire hence represents a “footprint” of the conditions faced by T cells that dynamically evolves according to the challenges that arise for the immune system, such as tumor neo-antigenic load. Hence, TCR repertoire analysis is becoming increasingly important to comprehensively understand the nature of a successful antitumor T-cell response, and to improve the success and safety of current CBI.

Nitric oxide reduction by carbon monoxide and carbon monoxide oxidation by O2 over Co–Mn composite oxide material

The promotional effect of precious metals in composite oxides has been observed in several catalytic reactions. In the present study, investigation of the CO oxidation with O2 and NO–CO redox reaction over Pd-containing cobalt–manganese composite was discussed. The composite oxides were prepared via glycine-aided combustion method. The physical properties of the catalysts were studied using X-ray diffraction, BET surface area, SEM, TEM and XPS. The strength of surface oxygen and surface reactivity of metal and CO were studied by H2-TPR and CO-pulse titration. It was observed that CO oxidation in cobalt–manganese composite oxide containing 5% PdO starts from 30 °C and the reaction gets completed at 65 °C. Also in NO–CO redox reaction, it shows complete detoxification at 170 °C with a good stability.

Inhibition of blood group antibodies by soluble substances

The presence of multiple alloantibodies or an antibody to a highprevalance antigen in a patient sample can pose challenges in antibody identification. The pattern of reactivity seen on an antibody panel may show various strengths of reactivity by different methods of testing or same strength of reactivity at one or more phases of testing. To ensure proper identification, multiple investigative tools may be used. We review one of these methods-inhibition by soluble substances-which has become an expansion of our toolbox within the past 10 years. Alloantibodies can be inhibited using specific soluble substances. These soluble substances occur naturally in various fluids or can be manufactured. When a patient sample contains multiple antibodies, clinically significant or not, inhibition of one may help determine specificities of others. Specific inhibition of a particular antibody will also help to confirm its presence.

Optimal Mean Arterial Blood Pressure in Extremely Preterm Infants within the First 24 Hours of Life

To define levels of mean arterial blood pressure (MABP) where cerebrovascular reactivity is strongest (MABPOPT) during the early transitional circulation in extremely preterm infants and to investigate the association between deviations above and below MABPOPT with intraventricular hemorrhage (IVH) and mortality. A total of 44 infants born at a median gestational age 25 (23-27) weeks with indwelling arterial catheter were studied at a median 5.5 (3.1-12.6) hours within the first 24 hours of life. Cerebrovascular reactivity (tissue oxygenation heart rate reactivity index) was estimated by the moving correlation coefficient between heart rate and near-infrared spectroscopy tissue oxygenation index. MABPOPT was defined as the MABP where tissue oxygenation heart rate reactivity index reaches minimum value. Deviations below or above MABPOPT values were calculated along with MABPOPT values during retrospective data analysis. MABPOPT was detected in all infants. The mean (SD) MABPOPT was 31.3 (±4.7) mm Hg. MABPOPT increased with increasing gestational age, R = 0.424; P = .004. Deviations below MABPOPT were greater in the IVH group (mean 2.7 mm Hg; 95% CI 2.0-3.5) compared with no-IVH (mean 1.7 mm Hg; 1.1-2.2), P = .006. In infants who died, the deviation below MABPOPT was greater (mean 3.3; 95% CI 1.9-4.8) compared with those who survived (mean 1.9 mm Hg; 95% CI 1.4-2.3), P = .015. Defining optimal MABP based on the strength of cerebrovascular reactivity within the first 24 hours of life is feasible and can provide an individualized approach to the care of extremely preterm infants. Deviations below MABPOPT were significantly associated with IVH and death.

Self-Reactivity and T-Cell Receptor Repertoire of Effector and Regulatory T Cells in Autoimmune Diabetes

T cell infiltration and accumulation in pancreatic islets during autoimmune diabetes is to a large degree driven by TCR reactivity to beta cell antigens. However, it is not fully clear whether the strength of TCR reactivity to beta cell antigens is an important determinant of effector or regulatory T cell function. We observed that CD5 expression is proportional to the level of self-reactivity based on Nur77-GFP reporter of TCR signaling and insulin tetramer staining of islet infiltrating T cells. Using RNAseq profiling, TCR sequencing, and cell transfer we show that CD5 segregates functionally and transcriptionally distinct effector and regulatory T cells, demonstrating that reactivity to beta cell antigens is an important determinant of effector and regulatory T cell function in autoimmune diabetes. Disclosure Y. Kong: None. M.L. Sprouse: None. Y. Jing: None. M. Bettini: None. M. Bettini: None.

Improving the performance of virtual crossmatch results by correlating with nationally-performed physical crossmatches: Obtaining additional value from proficiency testing activities

PurposeWhen donor specific HLA antibodies (DSA) are identified, the predictive value of whether a certain strength of reactivity (mean fluorescence intensity, MFI) leads to a positive crossmatch is uncertain. To determine this, we compared the DSA MFI results we generated locally for nationally distributed proficiency samples against the percentage of other laboratories reporting a positive crossmatch. MethodDSA MFI from single antigen beads reported by our laboratory for nationally-distributed proficiency testing survey samples was compared against the aggregate percentage of participating laboratories reporting the crossmatch positive using direct, antiglobulin-enhanced microcytotoxic (CDC-AHG), or flow cytometric methods from 2011 to 2015. Results180 surveys were analyzed. Positive CDC-AHG and flow cytometric crossmatches were associated with MFI greater than 8554 and 2748 respectively for HLA class I, and 6919 and 3707 respectively for class II. Institutional MFI less than 3000 had high positive predictive values (0.98, 0.85, 0.81) for negative direct, AHG, and flow crossmatches, while MFI greater than 8000 had high negative predictive values for a positive direct, AHG, and flow crossmatches (1.00, 1.00, 0.97). ConclusionReview of locally-generated MFI results as part of participating in proficiency testing allow for predictability of crossmatch results against other laboratories, providing a replicable model for other participating centers.

Level of self-reactivity modulates functional potential of effector T cells in autoimmune diabetes.

Abstract T cell infiltration and accumulation in pancreatic islets during autoimmune diabetes is largely dependent on TCR specificity for beta cell antigens. However, it is not fully clear whether the level of self-reactivity is an important determinant of T cell function in autoimmunity. We observed that CD5 expression is proportional to the level of self-reactivity based on Nur77-GFP reporter of TCR signaling and insulin tetramer staining of islet infiltrating T cells. Using RNAseq profiling, TCR sequencing, and cell transfer diabetes model we show that CD5 segregates functionally and transcriptionally distinct effector T cells. Transfer of CD5hi islet infiltrating T cells resulted in accelerated diabetes development compared to CD5lo cells (8 weeks vs. 11 weeks post transfer), suggesting that both T cell populations are able to re-infiltrate the pancreas and cause beta cell destruction, but CD5hi T cells exhibit increased pathogenicity. RNAseq analysis showed an increase in expression of Lag3 and FR4 in CD5hi cells, consistent with their increased activation and subsequent induction of regulatory and tolerogenic pathways. However, cell intrinsic negative regulators did not suppress IFNg production in CD5hi T cells, which was significantly higher compared to CD5lo cells. Our findings show that the strength of TCR reactivity to beta cell antigens is an important determinant of T cell pathogenicity in autoimmune diabetes.

Characterization of a hybrid protein designed with segments of allergens from Blomia tropicalis and Dermatophagoides pteronyssinus

BackgroundSensitization to allergens of the house dust mites Dermatophagoides pteronyssinnus and Blomia tropicalis is an important risk factor for asthma and allergic diseases. Allergen-specific immunotherapy is currently based on natural allergen extracts, however, in the last years recombinant allergens with different modifications have shown promising immunological properties that may be advantageously applied for developing novel allergy vaccines. MethodsA hybrid molecule (MAVAC-BD-2) containing epitopes of B. tropicalis (Blo t 5, Blo t 8 and Blo t 10) and D. pteronyssinus (Der p 1, Der p 2, Der p 7 and Der p 8) allergens was constructed, expressed in Escherichia coli and purified by affinity chromatography. Its folding was analyzed by circular dichroism. Antibody reactivities were evaluated by ELISA and non-denaturing dot blot assays using a battery of sera from mite allergic patients and non-allergic subjects. ELISA inhibition and dot blot assays with monoclonal antibodies were used to detect B-cell epitopes. Human basophil activation and induction of IgG-blocking antibodies in mice immunized with the hybrid protein were also evaluated. ResultsMAVAC-BD-2, expressed as a 22.8 kDa protein, showed a lower frequency and strength of IgE reactivity compared to Blo t 5, Der p 1, Der p 2 and the extracts of B. tropicalis and D. pteronyssinus. MAVAC-BD-2 inhibited 26% of IgE reactivity to Der p 2 and Blo t 5, reacted with anti-Der p 1 and anti-Der p 2 monoclonal antibodies and did not induce relevant basophil activation. MAVAC-BD-2 immunized mice produced specific antibodies that reacted against mite extracts and the purified allergens, as well as IgG antibodies that blocked the human IgE reactivity to mite extracts. ConclusionMAVAC-BD-2 has hypoallergenic characteristics and in mice induces IgG antibodies that block the human IgE reactivity to mite extracts.

A Subset of Human Autoreactive CD1c-Restricted T Cells Preferentially Expresses TRBV4-1+ TCRs.

In humans, a substantial portion of T cells recognize lipids presented by the monomorphic CD1 proteins. Recent studies have revealed the molecular basis of mycobacterial lipid recognition by CD1c-restricted T cells. Subsets of CD1c-restricted T cells recognize self-lipids in addition to foreign lipids, which may have implications in human diseases involving autoimmunity and malignancy. However, the molecular identity of these self-reactive T cells remains largely elusive. In this study, using a novel CD1c+ artificial APC (aAPC)-based system, we isolated human CD1c-restricted autoreactive T cells and characterized them at the molecular level. By using the human cell line K562, which is deficient in MHC class I/II and CD1 expression, we generated an aAPC expressing CD1c as the sole Ag-presenting molecule. When stimulated with this CD1c+ aAPC presenting endogenous lipids, a subpopulation of primary CD4+ T cells from multiple donors was consistently activated, as measured by CD154 upregulation and cytokine production in a CD1c-specific manner. These activated CD4+ T cells preferentially expressed TRBV4-1+ TCRs. Clonotypic analyses of the reconstituted TRBV4-1+ TCR genes confirmed CD1c-restricted autoreactivity of this repertoire, and the strength of CD1c reactivity was influenced by the diversity of CDR3β sequences. Finally, alanine scanning of CDR1 and CDR2 sequences of TRBV4-1 revealed two unique residues, Arg30 and Tyr51, as critical in conferring CD1c-restricted autoreactivity, thus elucidating the molecular basis of the observed V gene bias. These data provide new insights into the molecular identity of human autoreactive CD1c-restricted T cells.

The interaction of exaggerated and blunted reactivity with music on responses to unpleasant pictures.

Although personality can guide music listening habits (Rentfrow & Gosling, 2003), individuals use music for a multitude of purposes, many stemming from an aspect of emotion regulation in order to express or modify the experience of emotion (Juslin & Laukka, 2004). Both auditory and visual stimuli provide contextual cues that could be associated with emotions. The emotions evoked by the stimuli could in turn influence physiological reactions to stressful events (Lovallo, 2005b). Research has suggested the maladaptiveness of excessively strong or excessively muted physiological responses to stressful stimuli in the environment (Lovallo, 2011). Music provides a useful context under which to examine reactivity to stress, given that it has effects on the brain that lead to effects on peripheral functioning (Koelsch et al., 2007; Panksepp & Bernatzky, 2002). The current research aim was to explore individual differences in the strength of physiological and emotional reactivity in response to a stressor, namely unpleasant images, and whether the auditory soundscape could influence the level of reactivity.Models of emotion have been used to examine the relationship between music, emotion, and cardiovascular activity. The circumplex model, expounded by Russell (1980), denotes two aspects of emotion that are characterized as pleasantness and arousal. These two independent elements are postulated to intersect to form a circular emotion space comprised of four quadrants that contain differing types of emotions based on the interactions of arousal and pleasantness. Along the pleasantness dimension, individuals may experience varying levels of pleasantness or unpleasantness; along the arousal dimension individuals may experience varying levels of arousal. The intersection between arousal and pleasantness is thought to be derived from neurological activity that forms a prototypical core affect that persons can then use to infer their own emotional states, based on the surrounding social environment (Oatley & Johnson-Laird, 2014; Russell, 2003). Bradley, Greenwald, Petry, and Lang (1992) reported that memory of images one year after study was better if images were highly arousing thereby demonstrating that arousal is the more influential dimension of the two. Pleasantness can be related to motivational direction, such as approach or aversion, and arousal can be related to intensity or activation in the body. Another widely used approach to examining music and emotion has been the discrete model wherein emotions are discrete categories, such as happiness, anger, or sadness (Ekman, 1992). Here, emotions are universal, short lasting occurrences that have coherence between expression and autonomic output. We used the circumplex model in the current study on the basis of Eerola and Vuoskoski's (2011) contention that the dimensional nature of the model may provide advantages for consistency in emotion ratings compared with the discrete model.Research has investigated the effects of music elements on physiological and emotional responses. Tempo and mode, or the key of a music piece, are two widely studied elements. Research has found that faster tempo and the major mode are related to higher levels of physiological arousal, as assessed through cardiovascular measures such as heart rate and heart rate variability (Gomez & Danuser, 2007), although the relationship between mode and physiological responding is less corroborated (van der Zwaag, Westerink, & van den Broek, 2011). Mode, rhythmic articulation, and harmonic complexity contribute the most in distinguishing music as positively or negatively valenced (i.e., pleasant or unpleasant, respectively), whereas tempo, rhythmic articulation, and accentuation contribute in distinguishing music as high or low arousal (Gomez & Danuser, 2007) when applying twodimensional models. Unpleasant music is often written in the minor mode in contrast to the major mode (more often consonant) often heard in pleasant music (e. …

Weakly self-reactive T-cell clones can homeostatically expand when present at low numbers.

T-cell division is central to maintaining a stable T-cell pool in adults. It also enables T-cell expansion in neonates, and after depletion by chemotherapy, bone marrow transplantation, or infection. The same signals required for T-cell survival in lymphoreplete settings, IL-7 and T-cell receptor (TCR) interactions with self-peptide MHC (pMHC), induce division when T-cell numbers are low. The strength of reactivity for self-pMHC has been shown to correlate with the capacity of T cells to undergo lymphopenia-induced proliferation (LIP), in that weakly self-reactive T cells are unable to divide, implying that T-cell reconstitution would significantly skew the TCR repertoire toward TCRs with greater self-reactivity and thus compromise T-cell diversity. Here, we show that while CD4+ T cells with low self-pMHC reactivity experience more intense competition, they are able to divide when present at low enough cell numbers. Thus, at physiological precursor frequencies CD4+ T cells with low self-pMHC reactivity are able to contribute to the reconstitution of the T-cell pool.

Carbohydrate supplementation does not blunt the prolonged exercise-induced reduction of in vivo immunity.

Carbohydrate (CHO) supplementation during prolonged exercise is widely acknowledged to blunt in vitro immunoendocrine responses, but no study has investigated in vivo immunity. To determine the effect of CHO supplementation during prolonged exercise on in vivo immune induction using experimental contact hypersensitivity with the novel antigen diphenylcyclopropenone (DPCP). In a double-blind design, 32 subjects were randomly assigned to 120min of treadmill exercise at 60% [Formula: see text] with CHO (Ex-CHO) or placebo (Ex-PLA) supplementation. Responses were also compared to 16 resting control (CON) subjects from a previous study (for additional comparison with a resting non-exercise condition). Standardised diets (24h pre-trial) and breakfasts (3.5h pre-trial) were provided. Subjects received a primary DPCP exposure (sensitisation) 20min after trial completion, and exactly 28days later the strength of immune reactivity was quantified by magnitude of the cutaneous response (skin-fold thickness and erythema) to a low dose-series DPCP challenge. Stress hormones and leucocyte trafficking were also monitored. CHO supplementation blunted the cortisol and leucocyte trafficking responses, but there was no difference (P>0.05) between Ex-CHO and Ex-PLA in the in vivo immune responses (e.g. both ~46% lower than CON for skin-fold response). CHO supplementation does not influence the decrease in in vivo immunity seen after prolonged exercise. The effects with more stressful (or fasted) exercise remain to be determined. However, there appears to be no benefit under the conditions of the present study, which have practical relevance to what many athletes do in training or competition.

Site of reactivity models predict molecular reactivity of diverse chemicals with glutathione.

Drug toxicity is often caused by electrophilic reactive metabolites that covalently bind to proteins. Consequently, the quantitative strength of a molecule's reactivity with glutathione (GSH) is a frequently used indicator of its toxicity. Through cysteine, GSH (and proteins) scavenges reactive molecules to form conjugates in the body. GSH conjugates to specific atoms in reactive molecules: their sites of reactivity. The value of knowing a molecule's sites of reactivity is unexplored in the literature. This study tests the value of site of reactivity data that identifies the atoms within 1213 reactive molecules that conjugate to GSH and builds models to predict molecular reactivity with glutathione. An algorithm originally written to model sites of cytochrome P450 metabolism (called XenoSite) finds clear patterns in molecular structure that identify sites of reactivity within reactive molecules with 90.8% accuracy and separate reactive and unreactive molecules with 80.6% accuracy. Furthermore, the model output strongly correlates with quantitative GSH reactivity data in chemically diverse, external data sets. Site of reactivity data is nearly unstudied in the literature prior to our efforts, yet it contains a strong signal for reactivity that can be utilized to more accurately predict molecule reactivity and, eventually, toxicity.

Abstract C10: Immunoseroproteomic profiling of autoantibodies to tumor-associated autoantigens in African American and Caucasian men with prostate cancer

Abstract Cancer-associated serum autoantibodies are generally considered as “sensors” of molecular events associated with tumorigenesis, and thus constitute promising biomarkers for cancer detection and management, as well as tools for the identification of novel oncoproteins. Immunoseroproteomics is a standard approach for the identification of serum autoantibodies reacting with tumor-associated antigens (TAA) in human cancer populations. This approach entails using one- and two-dimensional gel electrophoresis of proteins from aggressive PCa cell lines, coupled with immunoblotting and mass spectrometry, to characterize the anti-TAA antibody repertoire in a given patient. Immunoseroproteomics has been applied mostly to cancer patients of European and Asian ancestry, and has not been used in African American (AA) patients with PCa. AA men show a disproportionately high incidence and mortality of PCa, compared to other ethnic groups, and display more aggressive tumors developing at a younger age. Recent epidemiological studies suggest that after adjusting for socioeconomic and health care-related factors, these disparities still persist, pointing to biological factors as contributors to these disparities. Growing evidence indicates that genes associated with immune function and autoimmunity are dysregulated in prostate tumors from AA men compared to Caucasian (CC) men. This prompted us to initiate a comprehensive analysis and comparison of the anti-TAA autoantibody response in AA and CC men with PCa by immunoproteomics. We hypothesized that this immune response might be augmented in AA patients with PCa, and that it could be used to identify novel blood biomarkers for early screening and management of the disease in AA men. In initial studies, we screened by immunoblotting of aggressive PCa cell lysates randomly selected serum samples from AA (n=10) and CC (n=13) men) with PCa. The initial results indicated that the autoantibody response in the AA cohort was much more robust in terms of autoantibody frequency and strength of reactivity than the response in CC patients. We used selected AA sera to identify novel TAA by immunoseroproteomics. Our initial analysis identified alpha enolase, fumarate hydratase, and annexin A1 as candidate TAAs in PCa. These proteins have been implicated in PCa in previous studies. Additional validation will be achieved by determining the frequency of the autoantibody response and the expression of these and other identified TAAs in PCa tissue microarrays. We are currently expanding our immunoseroproteomics analysis to sera from larger cohorts of AA and CC men (n&amp;gt;200 per group) with and without PCa, stratified by age and cancer stage. Our immunoproteomics approach will aid in the customization of TAA arrays for autoantibody profiling, leading to the identification of promising biomarkers for early PCa detection and monitoring tumor progression in the AA population. This approach may also reveal immune determinants underlying PCa health disparities. Citation Format: Tino W. Sanchez, Saied Mirshahidi, Nathan R. Wall, Colwick Wilson, Susanne Montgomery, Carlos A. Casiano. Immunoseroproteomic profiling of autoantibodies to tumor-associated autoantigens in African American and Caucasian men with prostate cancer. [abstract]. In: Proceedings of the Sixth AACR Conference: The Science of Cancer Health Disparities; Dec 6–9, 2013; Atlanta, GA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2014;23(11 Suppl):Abstract nr C10. doi:10.1158/1538-7755.DISP13-C10

A 13-question approach to resolving serological discrepancies in the transfusion medicine laboratory.

Laboratory professionals, consultants, and treating physicians may encounter discrepancies in serological testing results for numerous reasons; identifying the reason(s) for the presence of an unexpected antibody or antigen can be challenging. A question-based approach can be useful in identifying the underlying cause of the discrepancy. We describe a new approach to serological problems in a transfusion-service laboratory. The approach we outline herein is targeted towards a general transfusion medicine service, rather than a center that offers complex antibody investigations using specialized techniques. This question-based problem-solving approach considers patient factors including diagnosis, transfusion history, previous pregnancies, and medication history, along with serological test results: ABO and Rh groups, direct and indirect antiglobulin tests, reacting temperature of the antibody, effect of enzyme treatment of cells, strength of reactivity, and antibody reactivity with umbilical cord cells. We also demonstrate the usefulness of this approach through a case scenario.