Disease Severity in Patients Infected with Leishmania mexicana Relates to IL-1β

Leishmania mexicana can cause both localized (LCL) and diffuse (DCL) cutaneous leishmaniasis, yet little is known about factors regulating disease severity in these patients. We analyzed if the disease was associated with single nucleotide polymorphisms (SNPs) in IL-1β (−511), CXCL8 (−251) and/or the inhibitor IL-1RA (+2018) in 58 Mexican mestizo patients with LCL, 6 with DCL and 123 control cases. Additionally, we analyzed the in vitro production of IL-1β by monocytes, the expression of this cytokine in sera of these patients, as well as the tissue distribution of IL-1β and the number of parasites in lesions of LCL and DCL patients. Our results show a significant difference in the distribution of IL-1β (−511 C/T) genotypes between patients and controls (heterozygous OR), with respect to the reference group CC, which was estimated with a value of 3.23, 95% CI = (1.2, 8.7) and p-value = 0.0167), indicating that IL-1β (−511 C/T) represents a variable influencing the risk to develop the disease in patients infected with Leishmania mexicana. Additionally, an increased in vitro production of IL-1β by monocytes and an increased serum expression of the cytokine correlated with the severity of the disease, since it was significantly higher in DCL patients heavily infected with Leishmania mexicana. The distribution of IL-1β in lesions also varied according to the number of parasites harbored in the tissues: in heavily infected LCL patients and in all DCL patients, the cytokine was scattered diffusely throughout the lesion. In contrast, in LCL patients with lower numbers of parasites in the lesions, IL-1β was confined to the cells. These data suggest that IL-1β possibly is a key player determining the severity of the disease in DCL patients. The analysis of polymorphisms in CXCL8 and IL-1RA showed no differences between patients with different disease severities or between patients and controls

Statistical analysis of IL-1β −511 SNP.

<p>Statistical analysis of IL-1β −511 SNP.</p

IL-1β expression in serum of patients with cutaneous leishmaniasis.

<p>(A) Western blot of mature IL-1β (17 kDa) in 4 controls (1–4), 9 LCL patients (5–13) and 7 DCL patients (14–20). (B) Graph of percentage intensity of IL-1β bands with statistically significant differences between controls <i>vs</i> LCL patients, controls <i>vs</i> DCL patients and LCL <i>vs</i> DCL patients. (Identical symbols above different bars are statistically significant: p<0.05).</p

Analysis of IL-1β cytokine production by non-stimulated monocytes purified with anti-CD14 magnetic beads from PBMC of control subjects and patients with cutaneous leishmaniasis after 18 hours <i>in vitro</i> culture.

<p>Mean values obtained from 7 healthy control subjects and 5 patients with leishmaniasis. (Identical symbols above two different bars are statistically significant: p<0.05).</p

Immunohistochemistry for IL-1β and <i>Leishmania mexicana</i> staining in lesions of patients with cutaneous leishmaniasis.

<p>(A) IL-1β staining on cells of LCL patient with single small lesion; (B) small clusters of <i>Leishmania</i> parasites in LCL patient with small ulcer; (C) diffuse distribution of IL-1β in LCL patient with abundant ulcers; (D) disintegrated <i>Leishmania</i> in LCL patient with abundant ulcers; (E) diffuse distribution of IL-1β in DCL patient; (F) clusters with abundant intact <i>Leishmania</i> parasites in DCL patient. Red arrows show IL-1β⁺ staining and black arrows show <i>L. mexicana</i> staining. (G) Normal skin was used as negative control for IL-1β immunostaining. (H) Control staining with secondary antibody. All sections were counterstained with haematoxylin. (A–H) scale bar = 50 µm. Immunostaining in tissue sections was visualized at a magnification of 400×. We show a representative result of different types of lesions within each group: LCL patients with one small ulcer: (n = 8 for IL-1β staining and n = 17 for <i>L. mexicana</i> staining) (A and B); LCL patients with various ulcers (n = 3) (C and D); DCL patients (n = 6) (E and F).</p

Strategies and performance of the CMS silicon tracker alignment during LHC Run 2

The strategies for and the performance of the CMS silicon tracking system alignment during the 2015–2018 data-taking period of the LHC are described. The alignment procedures during and after data taking are explained. Alignment scenarios are also derived for use in the simulation of the detector response. Systematic effects, related to intrinsic symmetries of the alignment task or to external constraints, are discussed and illustrated for different scenarios