Relationship between serologic profile (ANCA type) and clinical features of renal involvement in ANCA-associated vasculitides.

AIM: To compare the frequency, clinical features and outcomes of renal involvement in ANCA-associated vasculitides (AAV) in patients with antibodies against proteinase-3 (pr3-ANCA) and myeloperoxidase (MPO-ANCA). MATERIALS AND METHODS: In our retrospective study we enrolled 264 patients, 94 males and 170 females, median age 53 [36; 62] years. Among them 157 were pr3-ANCA positive and 107 were MPO-ANCA positive. AAV was diagnosed according to ACR criteria and Chapel Hill consensus conference definition (2012). Median follow up was 44 [18; 93] months. We assessed baseline BVAS and VDI by the end of the follow up. Serum creatinine (sCr), estimated glomerular filtration rate (eGFR), hematuria and daily proteinuria were estimated. Diagnosis and stage of chronic kidney disease (CKD) and acute kidney injury (AKI) were established according to KDIGO guidelines (2012) and Scientific Society of Russian Nephrologists (2016). RESULTS: Renal involvement was present in 181 (68.6%) patients, and its frequency was similar in pr3-ANCA and MPO-ANCA subgroups. Patients with MPO-ANCA developed rapidly progressive glomerulonephritis and hypertension significantly more often than patients with pr3-ANCA: 50.7% vs 35.6% (p=0.049) and 46.1% vs 29.8% (p=0.029) respectively. At disease onset, median sCr was significantly higher and eGFR was significantly lower in patients with MPO-ANCA (p<0.05). 1-year and 5-year renal survival rates were similar in pr3-ANCA-positive (93.9% and 87.4% respectively) and MPO-ANCA positive patients (87.4% and 83.1% respectively). Median BVAS and VDI scores were significantly higher in pr3-ANCA subgroup. The number of patients who developed AAV relapse during 1-year follow up was also significantly higher in pr3-ANCA subgroup. The frequency of eye and ENT involvement was significantly higher in pr3-ANCA positive patients than in MPO-ANCA-positive patients. CONCLUSION: The frequency of extrarenal manifestations, clinical features of renal involvement and relapse rate are associated with AAV serotype.

[The frequency of ophthalmologic manifestations of granulomatosis with polyangiitis (Wegener's) and their relationship to systemic diseases]. / Chastota oftal'mologicheskikh proiavlenii granulematoza s poliangiitom (Vegenera) i ikh sviaz' s sistemnoi patologiei.

AIM: To estimate the frequency of lesions in the organ of vision in granulomatosis with polyangiitis (GPA) (Wegener's) and to determine their relationship to systemic diseases. SUBJECTS AND METHODS: The retrospective study enrolled 218 patients followed up at the E.M. Tareyev Clinic of Nephrology, Internal and Occupational Diseases, with a diagnosis of GPA. The frequency and association of ophthalmic manifestations with systemic involvement were statistically analyzed using PASW Statistics 18. RESULTS: The organ of vision was impaired in 48.1% of the patients with GPA. The most common manifestations were orbital space-occupying lesion (22.9%), conjunctivitis/episcleritis (14.7%), dacryocystitis (6.0%), and scleritis (4.6%). Orbital space-occupying lesions occurred more frequently in the local type of the disease (p=0.0003), and, on the contrary, the involvement of the conjunctiva and eyeball was seen in patients with the systemic types of GPA (p=0.02). CONCLUSION: The findings may suggest that the orbital lesion is an independent manifestation of GPA, which develops more commonly in its local type. Conjunctivitis/episcleritis is, on the contrary, more frequently seen in the active phase of the disease and generally in the involvement of other organs and systems.

Generation of high-intensity spectral supercontinuum of more than two octaves in a water jet.

In this paper, we demonstrate experimentally for the first time (to our knowledge) the generation of spectral supercontinuum (SC) of more than two octaves with high intensity in a water jet. The spectrum of the generated SC extends from 350 to 1400 nm, with intensities up to 1011 W/cm2, and its generation efficiency is more than 50%. For the pump intensity 3.0×1012 W/cm2 in the spectral range from 400 to 800 nm, the spectrum is nearly flat (less than 40% deviation), which is useful for many applications.

The Study of Serum Level and Immunochemical Properties of Natural Antibodies to N-Acetylglucosaminyl-N-Acetylmurarnyl Dipeptide in Healthy Donors.

ELISA assay showed that sera from each of 729 healthy donors contained antibodies to the minimal component of bacterial cell walls, N-acetylglucosaminyl-N-acetylmuramyt dipeptide (GMDP). Anti-GMDP antibody levels were determined for 686 sera, which were classified into 3 groups: high- (17.6%), medium- (68.7%) and low-responder (13.7%). Inhibition analysis performed on representative sera showed that a proportion contained specific anti-GMDP antibodies reacting only with GMDP (i.e., GMDP interaction with anti-GMDP antibodies was inhibited by GMDP only) whereas the remaining sera reacted both with GMDP and with tetrasaccharide (GlcNAc-MurNAc)(2) (i.e., GMDP interaction with anti-GMDP antibodies in the latter sera was inhibited by both GMDP and tetrasaccharide). Inhibition analysis indicated, moreover, that the anti-GMDP antibodies contained in high-responder sera had higher affinity than those present in low-responder ones: GMDP inhibited the GMDP + anti-GMDP antibody interaction by 88.7% in the former sera vs. 53% in the latter. Sera contained both IgM and IgG antibodies to GMDP, but the mean level of anti-GMDP IgG antibodies in the high-responder sera was 30 times higher than in the low-responder ones.

[Antigenic structure of the Pro-rich region 536-566 of Bordetella pertussis pertactin (P.69). II. Analysis of peptide-specific antibodies]. / Antigennoe stroenie Pro-bogatogo uchastka 536-566 pertaktina (P.69) Bordetella pertussis. II. Analiz peptid-spetsifichnykh antitel.

New method of analysis of subpopulations of peptide-specific polyclonal antibodies (Abs) without their elimination was developed which does not require radiolabeling. B-epitopes were analyzed which were recognized by rabbit Abs in synthetic peptides KAPPAPKPAPQPGPQPP (17P), QPPQPQPEAPAPQP (14P), and GPQPPQPPQP (10P) from Pro-rich region 536-566 of Bordetella pertussis pertactin. Antigenic structure was studied by indirect and competitive ELISA, immunoaffinity chromatography, isoelectric focusing, and immunoblotting. Abs against all peptides recognize native pertactin. Abs against 14P recognize antigenic determinant in region QPEPAPQP and they comprise two main subpopulations of Abs. Abs against 10P interact with two overlapping B-epitopes GPQPPQPPQP and QPPQP. Abs against 17P recognize an epitope in region KAPPAPKPAPQ and two additional overlapping antigenic determinants PGPQPP and PQPP. Minimal sequence recognized by Abs against P17 is PQPP but not QPP. Abs against 17P comprise approximately 20 major subpopulations; six or seven of these subpopulations effectively interact with the C-terminal region PGPQPP (6P).

[Antigenic structure of the Bordetella pertussis pertactin Pro-rich segment 536-566. I. Preparation and analysis of antisera to synthetic peptides]. / Antigennoe stroenie Pro-bogatogo uchastka 536-566 pertaktina Bordetellapertussis. I. Poluchenie i analiz antisyvorotok k sinteticheskim peptidam.

Rabbit antibodies to four synthetic peptides from the Pro-rich sequence of B. pertussis P.69 have been obtained. Prior to immunization, peptides KAPPAPKPAPQPGPQPP (17P), QPPQPQPEAPAPQP (14P), GPQPPQPPQP (10P) and PGPQPP (6P) were conjugated with keyhole limpet hemocyanin (KLH). Immunoenzymatic analysis revealed the following antibody titers: 1:4,000,000 (17P), 1:64,000 (14P) and 1:256,000 (10P). Serum antibodies to KLH-6P did not react with 6P-coated plates but interacted with the hexapeptide in the ovalbumin-6P conjugate. The sera to KLH-14P effectively recognized glutaraldehyde-formed epitopes. Using indirect immunoassay and competitive assay, it was shown that antisera to KLH-17P, KLH-14P, KLH-10P and KLH-6P did not react with unrelated antigens. Sera and affinity-purified antibodies to peptides 10P, 14P and 17P recognized native pertactin with variable efficiency.

The occurrence of natural antibodies to minimal component of bacterial cell wall (N-acetylglucosaminyl-N-acetylmuramyl dipeptide) in sera from healthy humans.

ELISA assay showed that sera from each of 729 healthy donors contained antibodies to the minimal component of bacterial cell walls, N-acetylglucosaminyl-N-acetylmuramyl dipeptide (GMDP). Anti-GMDP antibody levels were determined for 686 sera which were classified into 3 groups: high (17.6%), medium (68.7%), and low responder (13.7%). Inhibition analysis performed on representative sera showed that a proportion contained specific anti-GMDP antibodies reacting only with GMDP (i.e., GMDP interaction with anti-GMDP antibodies was inhibited by GMDP only) whereas the remaining sera reacted both with GMDP and with the tetrasaccharide (GlcNAc-MurNAc)2 (i.e., GMDP interaction with anti-GMDP antibodies in the latter sera was inhibited by both GMDP and the tetrasaccharide). Inhibition analysis indicated, moreover, that the anti-GMDP antibodies contained in high-responder sera had higher affinity than those present in low-responder ones: GMDP inhibited the GMDP + anti-GMDP antibody interaction by 88.7% in the former sera vs. 53% in the latter. Sera contained both IgM and IgG antibodies to GDMP, but the mean level of anti-GMDP IgG antibodies in the high-responder sera was 30 times higher than in the low-responder ones.

[Polypeptides from murine peritoneal macrophages recognizing glycosaminylmuramoyl dipeptide]. / Polipeptidy iz peritoneal'nykh makrofagov myshi, uznaiushchikh gliukozaminilmuramoildipeptidy.

By means of radioligand analysis, murine peritoneal macrophages were shown to express several hundreds cell surface high-affinity GMDP-binding sites with a binding constant 350 pM. Photoaffinity labeling followed by SDS-PAGE enabled us to identify inside these cells 32-34 and 38 kDa proteins, specifically binding GMDP. Proteins 32-34 kDa were also detected by Western blotting analysis using biotinylated conjugate of polyacrylamide with immobilized GMDP-Lys [(GMDP-Lys)-PAA-(Bi)] in cell lysate of murine peritoneal macrophages.

[Preparation and characteristics of monoclonal antibodies to N-acetylglucosaminyl-(beta1-4)-N-acetylmuramoyl-alanyl-D-isoglutamine]. / Poluchenie i kharakteristika monoklonal'nykh antitel k N-atsetilgliukozaminil-(beta1-4)-N-atsetilmuramoil-alanil-D izoglutaminu.

Hybridoma E6/1.2 was produced by fusion of splenocytes from mice immunized with N-acetylglucosaminyl-(beta 1-4)-N-acetylmuramyl-alanyl-D-isoglutamine (GMDP), conjugated to methylated BSA, with SP2/0 myeloma cells. GMDP-specific monoclonal antibody was IgG1 subtype and had affinity constant 2.10(9) M-1. According to competitive ELISA, the presence of the intact disaccharide fragment and the alanyl residue was critical for the GMDP-antibody interaction.

Muramyl peptide-binding sites are located inside target cells.

Using flow cytometry and fluorescence polarization analysis, specific muramyl peptide-binding sites were shown to be located inside T-lymphocytes, macrophages and neuroblastoma cells, but not inside B-cells. No binding sites were found on the cell surface. The number of binding sites for each cell type was determined. Two types of binding sites were observed for myelomonocytic WEHI-3 cells with Kd values of 21 and 540 nM. Inhibition analysis demonstrated that for effective binding, an intact glycopeptide molecule and D-configuration of isoglutamine residue are important.

[Structure-functional study of glycosaminylmuramoyl peptides. The effect of chemical modification of N-acetylglucosaminyl-N-acetylmuramoyldipeptide on its immunomodulating properties in vivo and in vitro]. / Strukturno-funktsional'noe issledovanie gliukozaminilmuramoilpeptidov. Vliianie khimicheskoi modifikatsii N-atsetilgliukozaminil-N-atsetilmuramoildipeptida na ego immunomoduliruiushchie svoistva in vivo i in vitro.

The structure-function relationships in a series of synthetic glucosaminylmuramyl peptides--glycopeptide adjuvants of the bacterial origin--have been investigated. Modification of the N-acetylglucosaminyl-(beta 1-4)-N-acetylmuramyl-L-alanyl-D-isoglutamine (GMDP) molecule affects its adjuvant and pyrogenic activity in vivo. GMDP is shown to readily stimulate the synthesis of the IgG2a and IgG3 of the immunoglobulin subclasses upon the secondary immune response to the protein antigen. The adjuvant effect correlates with the comitogenic test of the reaction of splenocytes blast-transformation in the presence of B-cell mitogen. No direct dependence has been revealed between the level of the glycopeptides adjuvant action and their effect on the IL-1 production by macrophages.

[Synthetic immunogenic complexes containing a peptide from the surface protein of the foot-and-mouth disease virus]. / Sinteticheskie immunogennye kompleksy na osnove peptida poverkhnostnogo belka virusa iashchera.

Synthetic constructions containing a peptide antigenic determinant (C-terminal peptide 205-213 of the surface VP1 protein of the foot-and-mouth disease virus, O1K strain), glucosaminylmuramayl dipeptide (GMDP), and polyionic synthetic carriers were prepared. The polymerized peptide and peptide-BSA conjugates were synthesized as well. Among the constructions obtained only peptide-BSA conjugate proved to be highly immunogenic. Application of synthetic constructions to design immunogenic complexes is discussed.