Detection of leucine-rich alpha-2-glycoprotein 1 containing immunocomplexes in the plasma of lung cancer patients with epitope-specific mAbs.

BACKGROUND: Lung cancer is the leading cause of cancer-related deaths worldwide. With the expectation of improved survival, tremendous efforts and resources have been invested in the discovery of specific biomarkers for early detection of the disease. Several investigators have reported the presence of cancer-associated autoantibodies in the plasma or serum of lung cancer patients. Previously, we used a monoclonal antibody (mAb) proteomics technology platform for the discovery of novel lung cancer-associated proteins. OBJECTIVE: The identification of specific protein epitopes associated with various cancers is a promising method in biomarker discovery. Here, in a preliminary study, we aimed to detect autoantibody-leucine-rich alpha-2-glycoprotein 1 (LRG1) immunocomplexes using epitope-specific monoclonal antibodies (mAbs). METHODS: We performed sandwich ELISA assays using the LRG1 epitope-specific capture mAbs, Bsi0352 and Bsi0392, and an IgG-specific polyclonal antibody coupled to a reporter system as the detection reagent. We tested the plasma of lung cancer patients and apparently healthy controls. RESULTS: Depending on the epitope specificity of the capture mAb, we were either unable to distinguish the control from LC-groups or showed a higher level of LRG1 and IgG autoantibody containing immunocomplexes in the plasma of non-small cell lung cancer and small cell lung cancer subgroups of lung cancer patients than in the plasma of control subjects. CONCLUSIONS: Our findings underline the importance of protein epitope-specific antibody targeted approaches in biomarker research, as this may increase the accuracy of previously described tests, which will need further validation in large clinical cohorts.

IL-7/αIL-7 mAb M25 immunocomplexes expand CD8+ T cells but paradoxically abrogate the antitumor activity of CTLA-4 and PD-1 blockage.

Supraphysiological levels of IL-7 induce increase counts of pre-B cells, naive T cells and memory phenotype CD8+ T cells. Immunocomplexes of IL-7 and αIL-7 mAb M25 (IL-7/M25) were described as IL-7 superagonist in vivo. Thus, treatment of mice with IL-7/M25 remarkably increases the size of the T cell pool. We decided to use IL-7/M25 in order to expand the T cell population prior to the administration of αCTLA-4 and αPD-1 mAbs in tumor-bearing mice and in turn boost the immunotherapy based on a combination of CTLA-4 and PD-1 blockage. We found that just four doses of IL-7/M25 increased the absolute numbers of splenocytes approximately fivefold and significantly shifted the CD4+:CD8+ T cell ratio in favor of CD8+ T cells. There was also a substantive increase in relative counts of memory phenotype CD8+ T cells (approximately threefold) within CD8+ T cells but a significant decrease (approximately 30%) in relative counts of Treg cells within CD4+ T cells. All these data suggest that IL-7/M25 offer a suitable approach to potentiate tumor immunotherapy through CTLA-4 and PD-1 blockage. Unexpectedly, IL-7/M25 significantly abrogated the antitumor activity of αCTLA-4 plus αPD-1 mAbs in the following mouse tumor models: MC-38 and CT26 colon carcinoma and B16F10 melanoma. This paradoxical effect of IL-7/M25 on the antitumor activity of CTLA-4 and PD-1 blockage was not mediated via either increased levels of IL-10 or TGF-ß in the sera or increased counts of IL-10-producing B or T cells in the spleen of mice injected with IL-7/M25. Thus, our work shows that caution should be exercised when combining two immunotherapy approaches together.

Inmunocomplejos circulantes en el lupus eritematoso sistémico / Circulating immunocomplexes in systemic erythematous lupus

Introducción: Los biomarcadores son claves en el diagnóstico y pronóstico del lupus eritematoso sistémico en el cual las manifestaciones clínicas son extremadamente complejas y heterogéneas. Objetivo: Determinar el valor clínico de los inmunocomplejos circulantes en pacientes con lupus eritematosos sistémico. Métodos: Se determinaron los niveles séricos de inmunocomplejos fijadores del C1q y de los anticuerpos anti-ácido desoxirribonucleico de doble cadena (anti-ADNdc), anti-nucleosoma (anti-Nuc) y anti-proteínas ribosomales (anti-RibP) por el ensayo inmuno-adsorbente ligado a enzima (ELISA) en 93 pacientes con diagnóstico de lupus eritematosos sistémico. Se utilizaron exámenes no paramétricos para probar la asociación entre los inmunocomplejos y los auto-anticuerpos. La frecuencia de nefritis lupica en los grupos de pacientes positivos y negativos de inmunocomplejos circulantes se comparó mediante Chi cuadrado. Resultados: Los inmunocomplejos se encontraron en 24 (25,8 por ciento) pacientes con lupus eritematosos sistémico. Los pacientes que presentaron los títulos más altos de inmunocomplejos fueron los positivos a los tres auto-anticuerpos usados (p=0,044). Se encontró correlación directa entre los niveles de anti-RibP y los inmunocomplejos (Rho=0,303, p=0,003) y entre los anti-ADNdc y anti-Nuc (Rho=0,449, p=0,001). La nefritis lúpica se presentó en 58,3 por ciento de pacientes con inmunocomplejos, y 31,9 por ciento pacientes negativos de inmunocomplejos (p=0,213). Conclusiones: Los inmunocomplejos circulantes caracterizaron una fracción menor de pacientes con lupus eritematosos sistémico. La presencia de estos no se asoció a los anticuerpos anti-ADNdc ni a la nefritis lupica(AU)

Introduction: Biomarkers are essential in the diagnosis and prognosis of systemic erythematous lupus in which clinical manifestations are extremely complex and heterogeneous. Objective: To determine the clinical value of circulating immunocomplexes in patients with systemic erythematous lupus. Methods: Serum levels of C1q-binding immunocomplexes and anti-double-stranded deoxyribonucleic acid (anti-dsDNA), anti-nucleosome (anti-Nuc) and anti-ribosomal proteins (anti-RibP) were determined by enzyme-linked immunosorbent assay (ELISA) in 93 patients diagnosed with systemic lupus erythematosus. Nonparametric tests were used to test the association between immunocomplexes and auto-antibodies. The frequency of lupus nephritis in the circulating immunocomplex positive and negative patient groups was compared using Chi square. Results: Immunocomplexes were found in 24 (25.8 percent) patients with systemic lupus erythematosus. The patients with the highest immunocomplex titers were positive for the three autoantibodies used (p = 0.044). A direct correlation was found between the levels of anti-RibP and immunocomplexes (Rho = 0.303, p = 0.003) and between anti-dsDNA and anti-Nuc (Rho = 0.449, p = 0.001). Lupus nephritis occurred in 58.3 percent of immunocomplex patients, and 31.9 percent immunocomplex negative patients (p = 0.213). Conclusions: Circulating immunocomplexes characterized a smaller fraction of patients with systemic lupus erythematosus. Their presence was not associated with anti-dsDNA antibodies or lupus nephritis(AU)

Vasculitis por reacción de hipersensibilidad tipo III / Vasculitis for type III hypersensitivity reaction

Los procesos inmunitarios son utilizados por el organismo para defenderse de la agresión de agentes infecciosos; no obstante, en ciertos casos, el organismo reacciona de forma inapropiada o excesiva ocasionando diversos tipos de daño tisular. Estas situaciones, que conocemos como hipersensibilidad, pueden tener aspectos positivos o negativos al poder causar ellos mismos la enfermedad. Se presenta el caso de una niña de 14 años de edad, que acude al Hospital Pediátrico Docente William Soler después de varios ingresos en otros centros de salud, donde se planteó el diagnóstico de un pie de madura. Después de varias investigaciones y con el antecedente de alergia a diferentes medicamentos, los cuadros de amigdalitis a repetición, los datos del laboratorio y la clínica que presentaba la paciente, se estableció el diagnóstico de una vasculitis por reacción de hipersensibilidad tipo III. Por las características tan atípicas del cuadro clínico de esta paciente y la dificultad para llegar a un diagnóstico es importante la presentación de este caso(AU)

The immune processes are used by the body to defend against the aggression of infectious agents; however, in certain cases, the body reacts inappropriately or excessively causing various types of tissue damage. These situations, which we know as hypersensitivity, can have positive or negative aspects by being able to cause the disease themselves. We present the case of a 14-year-old girl who attended the William Soler Pediatric Teaching Hospital after several admissions to other health centers, where the diagnosis of a mature foot was raised. After several investigations and with the history of allergy to different drugs, the recurrent tonsillitis symptoms, the laboratory data and the clinic presented by the patient, the diagnosis of a vasculitis due to type III hypersensitivity reaction was established. Because of the atypical characteristics of this patient's clinical picture and the difficulty in reaching a diagnosis, the presentation of this case is important(AU)

Vasculitis por reacción de hipersensibilidad tipo III / Vasculitis for type III hypersensitivity reaction

Los procesos inmunitarios son utilizados por el organismo para defenderse de la agresión de agentes infecciosos; no obstante, en ciertos casos, el organismo reacciona de forma inapropiada o excesiva ocasionando diversos tipos de daño tisular. Estas situaciones, que conocemos como hipersensibilidad, pueden tener aspectos positivos o negativos al poder causar ellos mismos la enfermedad. Se presenta el caso de una niña de 14 años de edad, que acude al Hospital Pediátrico Docente William Soler después de varios ingresos en otros centros de salud, donde se planteó el diagnóstico de un pie de madura. Después de varias investigaciones y con el antecedente de alergia a diferentes medicamentos, los cuadros de amigdalitis a repetición, los datos del laboratorio y la clínica que presentaba la paciente, se estableció el diagnóstico de una vasculitis por reacción de hipersensibilidad tipo III. Por las características tan atípicas del cuadro clínico de esta paciente y la dificultad para llegar a un diagnóstico es importante la presentación de este caso(AU)

The immune processes are used by the body to defend against the aggression of infectious agents; however, in certain cases, the body reacts inappropriately or excessively causing various types of tissue damage. These situations, which we know as hypersensitivity, can have positive or negative aspects by being able to cause the disease themselves. We present the case of a 14-year-old girl who attended the William Soler Pediatric Teaching Hospital after several admissions to other health centers, where the diagnosis of a mature foot was raised. After several investigations and with the history of allergy to different drugs, the recurrent tonsillitis symptoms, the laboratory data and the clinic presented by the patient, the diagnosis of a vasculitis due to type III hypersensitivity reaction was established. Because of the atypical characteristics of this patient's clinical picture and the difficulty in reaching a diagnosis, the presentation of this case is important(AU)

Reduction in the presence of cryoglobulins over time in the hemodialysis treatment.

BACKGROUND: The presence of cryoglobulins in patients with chronic kidney disease (CKD) on hemodialysis is well described. However, the generation of cryoglobulins during the dialysis treatment has yet to be established. The aim of the present study was to determine the presence of serum cryoglobulins over time in the dialysis treatment in patients with CKD not infected with hepatitis C virus (HCV). METHOD: Peripheral blood samples were collected at the beginning of dialysis treatment and at 30, 60, 90 and 120 days afterwards. Cryoglobulins were defined by the presence of immunocomplexes that precipitated in vitro with exposure to cold and resolubilized when rewarmed. The components of the cryoprecipitate were analyzed by radial immunodiffusion. RESULTS: In this study, 14 patients were included: 11 male and three female, aged 28-88 years, with mean time on hemodialysis of 57 ± 36 days at baseline. The presence of cryoglobulin, constituted by IgM, IgA, IgG and the C3 and C4 components of the complement, was observed in the serum of all patients at the beginning of hemodialysis. Sequence analyses showed that the amount of cryoprecipitate decreased during the dialysis treatment. CONCLUSION: There was a high prevalence of mixed cryoglobulins in CKD patients at the beginning of hemodialysis, and the amount of cryoprecipitate decreased during the treatment.

IL-2/anti-IL-2 mAb immunocomplexes: A renascence of IL-2 in cancer immunotherapy?

The in vivo biological activity of IL-2 can be dramatically increased by complexing with anti-IL-2 mAb. Moreover, IL-2/anti-IL-2 mAb immunocomplexes selectively stimulate different subsets of immune cells, depending on the clone of anti-IL-2 mAb that is used. Thus, IL-2/S4B6 mAb complexes strongly stimulate CD122high populations, namely NK and memory CD8+ T cells. They also intermediately stimulate Treg cells. Conversely, IL-2/JES6.1 mAb immunocomplexes have no stimulatory activity for CD122high populations. However, they potently and highly selectively stimulate CD25+ cells (i.e., Treg and activated T cells). IL-2/S4B6 mAb immunocomplexes have also been shown to possess antitumor activity in various mouse tumor models.

Squamous cell carcinoma antigen-IgM is associated with hepatocellular carcinoma in patients with cirrhosis: A prospective study.

BACKGROUND: Squamous cell carcinoma antigen (SCCA)-IgM complex has been described as a promising tool to identify patients with progressive liver disease at higher risk of hepatocellular carcinoma (HCC) development in retrospective studies. AIM: To assess the clinical value of this biomarker in patients with cirrhosis in a prospective study. METHODS: Patients with overt cirrhosis were prospectively evaluated at 6-month intervals for HCC development and decompensation with clinical examination, liver ultrasound, α-fetoprotein measurement. SCCA-IgM was measured in serum by immunoenzymatic assay. Median follow-up duration was 52 months (range 12-68 months). RESULTS: 70 patients (26% male; mean age 56±10 years) were enrolled. The main aetiological factors were alcohol (44%) and hepatitis C (34%). Baseline values of SCCA-IgM were significantly higher in patients who developed HCC. Positivity of the biomarker at baseline was associated with a significantly shorter HCC-free survival, while α-fetoprotein (cut off >20 ng/ml) was not significant. SCCA-IgM positivity and hepatitis C were significant prognostic factors for HCC development. The biomarker was not associated with the development of clinical complications of cirrhosis. CONCLUSION: This prospective study demonstrates that in patients with cirrhosis SCCA-IgM is associated with HCC development and may be useful for clinical management of cirrhotic patients at higher risk of HCC development.

Increasing the biological activity of IL-2 and IL-15 through complexing with anti-IL-2 mAbs and IL-15Rα-Fc chimera.

IL-2 and IL-15 are structurally relative cytokines that share two receptor subunits, CD132 (γ(c) chain) and CD122 (ß chain). However, the expression pattern and physiological role of IL-2 and IL-15 private receptor α chains CD25 and IL-15Rα, respectively, are strikingly different. CD25, together with CD122 and CD132, forms a trimeric high affinity IL-2 receptor that is expressed and functions on cells acquiring an IL-2 signal. Conversely, IL-15Rα is expressed and binds IL-15 with high affinity per se already in the endoplasmic reticulum of the IL-15 producing cells and it presents IL-15 to cells expressing CD122/CD132 dimeric receptor in trans. Thus, while IL-2 is secreted almost exclusively by activated T cells and acts as a free molecule, IL-15 is expressed mostly by myeloid cells and works as a cell surface-associated cytokine. Interestingly, the in vivo biological activity of IL-2 can be dramatically increased through complexing with certain anti-IL-2 mAbs; such IL-2/anti-IL-2 mAbs immunocomplexes selectively stimulate the proliferation of a distinct population of immune cells, depending on the clone of the anti-IL-2 mAb used. IL-2/S4B6 mAb immunocomplexes are highly stimulatory for CD122(high) populations (memory CD8(+) T and NK cells) and intermediately also for CD25(high) populations (Treg and activated T cells), while IL-2/JES6-1 mAb immunocomplexes enormously expand only CD25(high) cells. Although IL-2 immunocomplexes are much more potent than IL-2 in vivo, they show comparable to slightly lower activity in vitro. The in vivo biological activity of IL-15 can be dramatically increased through complexing with recombinant IL-15Rα-Fc chimera; however, IL-15/IL-15Rα-Fc complexes are significantly more potent than IL-15 both in vivo and in vitro. In this review we summarize and discuss the features and biological relevance of IL-2/anti-IL-2 mAbs and IL-15/IL-15Rα-Fc complexes, and try to foreshadow their potential in immunological research and immunotherapy.

Changes and significance of the circulating immunocomplexes in patients with recurrent uveitis / 眼科新进展

Objective　To explore the changes and significance of the circulating immunocomplexes(CIC) in patients with chronic iridocyclitis.Methods　We assessed circulating immunocomplexes in peripheral blood of 45 patients with recurrent uveitis and 42 healthy controls by PEG deposition with the biochemical analyzer to obtain levels of CIC different with recurrent uveitis.Results　Levels of CIC were significantly higher in active patients(27.358±6.729)than those in controls(13.459±6.164,P＜0.001) and inactive patients (16.248±6.233,P＜0.001),but patients (inactive) had significantly higher levels than those in controls (P＜0.05).Conclusion　Levels of CIC were found in the groups of active patients with recurrent uveitis (27.358±6.729), but levels of CIC in inactive patients fell. The result of this study suggest that recurrence of uveitis is related to immunocomplexes,and changes in the levels of CIC could be also a measure standard of therapeutic effects.