Heterozygous nucleotide-binding oligomerization domain-2 mutations affect monocyte maturation in Crohn's disease

AIM： To investigate the function of monocytes in Crohn＇s disease （CD） patients and to correlate this with disease- associated nucleotide-binding oligomerization domain-2 （NOD2） gene variants. METHODS： Monocytes from 47 consecutively referred CD patients and 9 healthy blood donors were cultured with interleukin （IL）-4 and granulocyte-macrophage colony-stimulating factor （GM-CSF）, and stimulated with lipopolysaccharide （LPS） or muramyldipeptide （MDP）, the putative ligand of NOD2. RESULTS： We found that monocytes from CD patients differentiated in vitro to mature dendritic cells （DCs）, as determined by immunophenotype and morphology. IVOD2 genotype was assessed in all subjects, and we observed high CD86 expression on immature and LPS-stimulated DCs in IVOD2 mutated CD patients, as compared with wtlVOD2 CD patients and controls. By contrast, CD86 expression levels of DCs induced to maturity with MDP derived from IVOD2-mutated subjects were comparable to those of normal subjects. The amount of IL-12p70 in patient-cell cultures was larger than in controls aEer LPS treatment, but not aEer treatment with MDR CONCLUSION： Our results suggest that DCs obtained from patients with mutations in the IVOD2 gene display an activated phenotype characterized by high CD86 expression, but have a diminished response to MDP when compared to the terminal differentiation phase. We speculate that the altered differentiation of monocytes might lead to an imbalance between inflammation and the killing ability of monocytes, and may be relevant to the pathogenesis of CD.

A new approach for the treatment of CLL using chlorambucil/hydroxychloroquine-loaded anti-CD20 nanoparticles

Current approaches for the treatment of chronic lymphocytic leukemia （CLL） have greatly improved the prognosis for survival, but some patients remain refractive to these therapeutic regimens. Hence, in addition to reducing the long-term side- effects of therapeutics for all leukemia patients, there is an urgent need for novel therapeutic strategies for difficult-to-treat leukemia cases. Due to the cytotoxicity of drugs, the major challenge currently is to deliver the therapeutic agents to neoplastic cells while preserving the viability of non-malignant ceils. In this study, we propose a therapeutic approach in which high doses of hydroxychloroquine and chlorambucil were loaded into biodegradable polymeric nanopartides coated with an anti-CD20 antibody.We first demonstrated the ability of the nanoparticles to target and internalize in tumor B-cells. Moreover, these nanoparticles could kill not only p53-mutated]deleted leukemia cells expressing a low amount of CD20, but also circulating primary ceils isolated from chronic lymphocytic leukemia patients. The safety of these nanoparticles was also demonstrated in healthy mice, and their therapeutic effects were shown in a new model of aggressive leukemia. These results showed that anti-CD20 nanoparticles containing hydroxychloroquine and chlorambucil can be effective in controlling aggressive leukemia and provided a rationale for adopting this approach for the treatment of other B-cell disorders.

ntestinal-mucosa anti-transglutaminase antibody assays to test for genetic gluten intolerance

Celiac disease （CD） is an autoimmune enteropathy character- ized by gluten-triggered intestinal mucosa lesions in genetically susceptible individuals carrying the HLA DQ2 or DQ8. CD diagnosis is based on the concentration of IgA serum anti- transglutaminase （anti-tTG） antibodies together with mucosal damage at intestinal biopsy.