Longitudinal Zeolite-Iron Oxide Nanocomposite Deposited Capacitance Biosensor for Interleukin-3 in Sepsis Detection

Sepsis is an extreme condition involving a physical response to severe microbial infection and causes fatal and life-threatening issues. Sepsis generates during the chemicals release with the immune system into the bloodstream for fighting against an infection, which causes the inflammation and leads to the medical emergency. A complexed longitudinal zeolite and iron oxide nanocomposite was extracted from coal mine fly ash and utilized to improve the surface characteristics of the capacitance biosensor to identify sepsis attacks. Anti-interleukin-3 (anti-IL-3) antibody was attached to the zeolite- and iron oxide-complexed capacitance electrode surface through an amine linker to interact with the sepsis biomarker IL-3. The morphological and chemical components of the nanocomplex were investigated by FESEM, FETEM, and EDX analyses. At approximately 30 nm, the longitudinal zeolite and iron oxide nanocomposite aided in attaining the limit of IL-3 detection of 3 pg/mL on the linear curve, with a regression coefficient (R2) of 0.9673 [y = 1.638x − 1.1847]. A lower detection limit was achieved in the dose-dependent range (3–100 pg/mL) due to the higher amount of antibody immobilization on the sensing surface due to the nanomaterials and the improved surface current. Furthermore, control experiments with relevant biomolecules did not show capacitance changes, and spiked IL-3 in human serum increased capacitance, indicating the specific and selective detection of IL-3. This study identifies and quantifies IL-3 via potentially useful methods and helps in diagnosing sepsis attack.

Purification of recombinant human interleukin-3 expressed as inclusion bodies in Escherichi coli

Human interleukin-3 (IL-3) is a hematopoietic growth factor involved in the survival, proliferation and differentiation of multipotent hematopoietic cells. However, recombinant IL-3 is usually expressed as insoluble form (inclusion bodies) in Escherichia coli cells. This state of protein often shows no bioactivity. Herein, we report a simple method for solubilization, refolding and purification of recombinant human IL-3 expressed in E. coli cells. First, IL-3 was expressed in E. coli JM109 (DE3) after being induced with 0.05 mM IPTG at 25 oC. Under these conditions, IL-3 was produced as inclusion bodies with molecular weight of approximately 15 kDa on SDS-PAGE gel (14%). Next, IL-3 pellet was separated from the host soluble proteins using sonication followed centrifugation. Then, two strong denaturants such as urea or guanidine hydrochloride were used to test solubilization of the insoluble IL-3. After that, the resulting soluble IL-3 was renatured and subjected to gel filtration chromatography to collect purified IL-3 protein. Our results showed that fractionates contained a single band of IL-3 with recovery rate of about 30%. Several characteristics of recombinant IL-3 were then analyzed. The cytokine IL-3 showed its high purity with a sharp peak on RP-HPLC chromatagram. The Western blot showed a clear signal band on PVDF membrane to demonstrate its right antigenecity against human IL-3 antibody. Besides, amino acid sequence of this cytokine was confirmed by mass spectrophotometry method. The purified IL-3 cytokine is a potential material for further tests.

Increased disease burden in Interleukin-3 deficient mice after Mycobacterium tuberculosis and herpes simplex virus infections

Interleukin-3 (IL-3) is produced during infections caused by parasites, bacteria and viruses, but its contribution to immunity in this context remains largely unknown. In mouse models of parasitic infections, in which the effects of IL-3 have been most extensively studied, IL-3 has been variously reported as protective, detrimental or inconsequential. Similarly, mixed results have been reported in viral and bacterial infection models. Here, we investigated the effects of IL-3 in mouse models of Mycobacterium tuberculosis and herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) infections by assessing the pathogen burden, disease manifestations and survival following infection. After infection with M. tuberculosis, IL-3 deficient mice showed higher bacillary burden, increased lung pathology and reduced survival compared to wild type mice. After infection with HSV-1 through cutaneous route and HSV-2 through vaginal route, IL-3 deficient mice showed higher viral burden, increased disease manifestations and reduced survival compared to wild type mice. Our results show that IL-3 makes a subtle but significant contribution to protective immunity in these mouse models of bacterial and viral infections.

The role of interleukin-3 and its receptor in acute leukemia pathogenesis

Introduction. Interleukin-3 (IL-3) is the key cytokine involved in the regulation of normal haematopoiesis. Some leukemic cells demonstrate high expression of the α-subunit of the receptor for interleukin-3 (CD123).Aim: to summarize the current understanding of IL-3 and its receptor CD123 in the pathogenesis of acute leukemia. General fi ndings: IL-3 regulates the proliferation and differentiation of normal hematopoietic progenitor cells in the early stages of hematopoiesis. The IL-3 receptor (CD123) is expressed on normal hematopoietic cells. High expression of CD123 was confi rmed on blast cells of AML, B-ALL and on the leukemia-initiating CD34+ CD38– cells. IL-3 inhibits apoptosis and promotes the autonomous growth of blast cells. Currently, different approaches of blocking the IL-3 mediated signal are being investigated.