Humán papillomavírusok szerepe a szájüreg daganatos elváltozásaiban = Human papillomaviruses in oral cancers

Magyarországon a szájüregi rosszindulatú daganatok előfordulása és mortalitása kiemelkedően magas. Célkitűzésünk a humán papillomavírus (HPV) fertőzés szájüregi tumorgenezisben játszott etiológiai szerepének és prognosztikai indikátorként való alkalmazhatóságának vizsgálata volt, emellett vizsgáltuk a p53 tumorszuppresszor gén 72 kodont érintő polimorfizmusának és mutációinak, valamint az 1. genocsoportú torque-tenovírus (g1TTV) prognosztikai szerepét is. Mintáink 65 orális laphámrákos (OSCC), 44 orális leukoplakiában és 119 orális lichen planusban szenvedő betegtől származtak, a kontroll populációt 72 egészséges személy képezte. A HPV prevalenciája a lézió súlyosságával nőtt, szignifikánsan magasabb volt minden betegcsoportban, mint a kontrollban, főképpen magas kockázatú genotípusokat találtunk. Emellett a HPV jelenléte a tumor lokalizációjával illetve kémiai karcinogénekkel (alkohol, dohányzás) együttesen hatva a OSCC rossz pronózisával és a betegek rosszabb túlélésével társult. A g1TTV és a p53 gén polimorfizmusa és mutációi nem befolyásolták az OSCC prognózisát. A projekt adatai alátámasztják a HPV etiológiai szerepét a OSCC kialakulásában, emellett azt mutatják, hogy a prekancerózisok kialakulásában is szerepet játszhat. Igazoltuk, hogy a HPV jelenléte kockázati tényezőként szerepel a rossz prognózisú tumorok kialakulása során, tehát a HPV kimutatása segítséget nyújthat a prognózis megállapításában és az utókezelés tervezésében. | Morbidity and mortality due to oral squamous cell carcinoma (OSCC) is high in Hungary. The aim of the project was to investigate the role of human papillomaviruses (HPVs) in the etiology of OSCC as well as its prognostic value. Besides HPVs we also examined the prognostic significance of genogroup 1 torque-tenovirus (g1TTV) and the mutations and codon 72 polymorphism of p53 tumour suppressor gene. Study population consisted of 65 OSCC, 44 oral leukoplakia and 119 oral lichen planus patients, buccal epithelial cells from 72 healthy individuals were used as a control. Prevalence of HPV increased gradually with the increasing severity of the lesion, it was significantly higher in all patients groups compared to the control. We detected mainly high-risk genotypes. Presence of HPV synergistically with unfavourable tumour localization and chemical carcinogens (alcohol consumption, smoking) led to higher risk of poor prognosis and shorter survival. Presence of g1TTV and p53 gene mutations and polymorphism did not later the prognosis significantly. Our data accord with etiologic role of HPVs in OSCC, and indicate that HPVs may play a role in the development of oral precanceroses as well. We proved that presence of HPV may serve as a risk factor for tumours with poor prognosis, therefore it can be used as a prognostic indicator and may aid in planning adjuvant therapy

Humán papillomavírus és humán herpeszvírusok kóroki szerepének molekuláris vizsgálata = Molecular analysis of pathogenic role of human papilloma virus and human herpesviruses

Vesetranszplantáltak mintáiban a humán herpeszvírus 6A dominanciáját, és a humán cytomegalovírussal megegyező gyakoriságát találtuk. Az új KI és WU polyomavírusok jelenlétét mi írtuk le először vizeletből, a HPyV9-et légúti mintákból. A WU és KI vírusokat vesetranszplantáltak légúti és vérmintáiban is kimutattuk, egészséges személyekben nem. A HPyV9-et vér és vizeletmintákból is kimutattuk. Humán papillomavírus 11 pozitív mintákból a vírus genomjának szekvenálását, metilációs mintázatát vizsgálva tanulmányoztuk a cidofovir terápia kudarca hátterében álló genetikai, epigenetikai változásokat. Nem mutattunk ki a vírus genomot érintő genetikai és epigenetikai változásokat. A virális LCR-ben (long control region) egyedi polimorfizmusok/mutációk magyarázhatják a vírus magas virulenciáját, ami hozzájárulhat a terápia kudarcához. Vizsgáltuk a torque teno vírus genetikai diverzitását a magyar populációban, a fej-nyaki régióból és a női genitális traktusból, normál, precancerosus és malignus léziók mintákban. Az összes TTV szekvencia a TTV1 faj típus-szekvenciához illeszkedett. A leggyakoribb szubtípus a 2c-volt. A 2b és a2c szubtípus egyenlő arányban fordult elő a cervicalis mintákban; az 1a szubtípus viszont gyakrabban fordul elő a cervicalis atypiában és méhnyakrákban. A fej-nyaki minták körében a 2c szubtípus volt a leggyakoribb. A szubtípusok megoszlása földrajzi régiónként és etnikumonként jelentős különbségeket mutat. | We found that A variant of human herpesvirus 6 is dominant in renal transplant patients and as frequent as the human cytomegolvirus proving that monitoring is important after the transplantation. Newly described human polyomavirus KI and WU were found and published first in urine. Theye were detected in respiratory and blood samples from renal transplant patients, but not healthy individuals nor pregnant women suggesting that these viruses might have importance mainly in immunocompromised patients. HPyV9 was found and published first in respiratory samples suggesting a new hypothesis about the transmission. The viruses was found also in urine and blood samples. Human papillomavirus type 11 positive samples were collected fand analyzed. Sequencing of the genome, methylation analysis were performed to seek for genetic and epigenetic changes as a possible background for therapy failure. We found that the virological failure was not due to virological factors suggesting that cidofovir action may depend more heavily on the host. Diversity of torque teno virus was assessed in the head and neck region in patients with potentially malignant and malignant lesions and was compared to that found in the uterine cervix (cervical atypia and cervical cancer) by sequencing. The results are that genotype and even subtype distribution may be important in association with diseases, therefore using this classification for characterization of intraspecies diversity of TTV1 is proposed

Humán papillomavírus fertőzések = Human papillomavirus infections

A survivin kifejeződése fontos lehet a méhnyakrák kialakulásában. A survivin és a HPV (human papillomavírus) közötti kapcsolatot vizsgálva megállapították, hogy a survivin promóter polimorfizmus nem jelent kockázati tényezőt a cervix carcinoma kialakulásában. Megállapították, hogy a HPV 16 E7 fehérjéje gátolja a TGF-?2 promóter transzkripciós aktivitását az E2F1 Rb/E2F komplexből történő felszabadítása révén. A gége laphámsejtes karcinomás betegekben, ha az 1. genocsoportú TTV (human circovírus)-fertőzés HPV fertőzéssel társul, szignifikánsan romlik a betegek tumorprogresszió-mentes túlélése. Az E cadherin ? 160 C/A polimorfizmusának megléte és a HPV jelenléte között nincs összefüggés a cervix carcinomás és a laryngeális carcinoma planocellulare daganatokból származó mintákban. A laryngeális papilloma planocellulare mintákban viszont szignifikánsan nagyobb volt a polimorf A allél frekvenciája, mint a kontroll populációban. Cidofovir terápiára a rekurrens respiratory papillomatosis (RRP) az első kéthetes kezelés alatt jelentősen javult, s a mindig kimutatható HPV-11 genom kópiaszám jelentősen lecsökkent. | Expression of survivin may play an important role in development of cervical carcinoma. Studying the interaction between survivin and HPV (human papillomavirus) it was suggested that promoter polymorphism of survivin gene is not a risk factor in cervical carcinoma. It was suggested that E7 protein of HPV 16 inhibits the transcriptional activity of TGF-?2 promoter by releasing E2F1 from Rb/E2F complex. If TTV (human circovirus) genogroup 1 infection is accompanied by HPV infection in patients with squamous cell carcinoma of the larynx progression free survival of patients are significantly decreased. Correlation between ? 160 C/A polymorphism of E cadherin and presence of HPV in samples originated from cervical cancer and carcinoma planocellulare of larynx was not detected. In case of laryngeal papilloma planocellulare the frequency of polymorph A allel was found significantly higher than in the control population. Owing to the cidofovir therapy recurrent respiratory papillomatosis (RRP) shows remarkable improvement, and the copy number of HPV 11 detected every time decreased considerably

Genital and oral carriage of human papillomaviruses in women with high grade squamous intraepithelial lesionand their male partners

Human papillomaviruses (HPVs) are the main risk factor for cancer of the uterine cervix, and may also play a role in rectal, penile as well as head and neck cancer. A major way of HPV transmission is sexual contact, oropharygeal HPVs may derive from oral sexual contact with genitally infected partners. The aim of the present work was to explore genital as well as oral HPV status in women with high grade squamous intraepithelial lesion (HSIL) and in their male partners. Thirty-threewomen (median age 27; range 19-59) with cytologically confirmed HSIL and their male partners (median age 31, range 23-65) were enrolled; informed consent was collected from all participants. Exfoliated cells from the oral and genital mucosa were collected by cytobrush and placed immediately in dry ice. All samples from the couples were collected within amonth to assess simultaneous carriage of HPVs. HPV detection was performed by MY/GP consensus nested PCR, HPVs were genotyped by sequencing and virus copy numbers were determined using real-time PCR. Prevalence data were compared by chi-square test or sign test, as appropriate. Log copy numbers were compared using Kruskal-Wallis test with Bonferroni correction. ANOVA and Yule’s coefficient was used to test for association of HPV status with that of the partner. As expected, the majority of women with HSIL proved to be HPV positive in the genital mucosa (28/33). Their oral mucosa, however, showed significantly lower HPV carriage (7/33, p<0.001); all orally positive women carried HPV in the genital mucosa. Similarly, males carried HPV in their genital mucosa more frequently than in the oral mucosa (17/33 vs. 7/33, p=0.035), but oral carriers were not always positive in their genital sample. Females carry HPV more frequently than males in their genital (p=0.004) but not in their oral samples. The most frequently encountered genotype was HPV16 in both sexes, but high risk genotypes HPV18, 31, 33, 51, 56, 66, 82 and low risk HPV11, 55, 61, 72, 81, 83 and 84 were also found. The genotype was the same in the genital and oral mucosa of 7/7 and 5/7 females and males, respectively. Average copy numbers in positive females were significantly higher in the genital than in the oral mucosa (1.3x105 vs 1.5x103/ug DNS, p=0.003), while in males the copy numbers were comparable (8.2x103 vs 1.8x102/ug DNS). Females had higher copy numbers than males in the genital (p<0.001), but not in the oral mucosa. There were no differences in copy numbers of females with HPV positive or negative partners. In couples, females are more frequently carriers than males in the genital (p<0.002), but not in the oral mucosa. In the genital mucosa 15, while in the oral mucosa three pairs shared the same HPV genotype, but neither genital nor oral carriage was found to be associated statistically with the status of the partner. Though transmission between oral and genital mucosa as well as transmission between sexual partners is plausible, proving this association statistically needs higher sample sizes with enrolment of more couples. This work was supported by a János Bolyai Research Scholarship of the Hungarian Academy of Sciences

Asymptomatic faecal carriage of ESBL producing Enterobacteriaceae in Hungarian healthy individuals and in long-term care applicants: a shift towards CTX-M producers in the community

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A WFIKKN fehérjék és a miosztatin, GDF11 közötti kölcsönhatás jellemzése = Characterization of the interaction of WFIKKN1 and WFIKKN2 with Myostatin and GDF11.

Kimutattuk, hogy a WFIKKN fehérjék specifikus és hatékony inhibitorai a miosztatin/GDF8 és GDF11 növekedési faktoroknak. - Felületi plazmon rezonancia (SPR) mérésekkel megállapítottuk, hogy mindkét fehérje nagy affinitással (Kd ~ 10-8- 10-10M) köti a GDF8 és GDF11 növekedési faktorokat. A WFIKKN fehérjék elsősorban a follisztatin homológ (FS) doménjük révén kötik a miosztatint, de a netrin homológ (NTR) domén is részt vesz a kölcsönhatás kialakításában. - Kompeticiós SPR mérésekkel kimutattuk, hogy a WFIKKN fehérjék hatékonyan gátolják a miosztatin és GDF11 kötődését a receptoruk, az Activin-Receptor IIb ligand-kötő doménjéhez. - A miosztatin és GDF11 jelátvitel detektálására alkalmas riporter rendszerben mindkét WFIKKN fehérje hatékonyan gátolta a miosztatin és GDF11 növekedési faktorok által kiváltott jelátvitelt. - SPR mérésekkel megállapítottuk, hogy a WFIKKN fehérjék jelentős affinitással (Kd ~ 10-7- 10-8M) kötnek több más, a TGF? családba tartozó növekedési faktort is: a TGF?1-t, BMP2-t, BMP4-t. Gyengébb kölcsönhatást detektáltunk a BMP3 és BMP8b-vel és nem volt mérhető kölcsönhatás az Aktivin A-val. - A WFIKKN fehérjék és a TGF?1, BMP2 és BMP4 növekedési faktorok közötti kölcsönhatás ellenére riporter vizsgálatokban a WFIKKN fehérjék nem gátolták a TGF?1, BMP2 és BMP4 által kváltott jelátvitelt. Összegezve: eredményeink azt mutatják, hogy a WFIKKN1 és WFIKKN2 a miosztatin és GDF11 növekedési faktorok specifikus inhibitorai. | We have shown that WFIKKN proteins are potent and specific inhibitors of myostatin/GDF8 and GDF11, since - surface plasmon resonance measurements revealed that both WFIKKN1 and WFIKKN2 have high affinity (Kd ~ 10-8- 10-10M) for myostatin/GDF8 and GDF11. The interactions are mediated primarily by the follistatin homolog domains of WFIKKN proteins, but their NTR domains also contribute to these protein-protein interactions. - competitive SPR measurements have shown that WFIKKN1 and WFIKKN2 inhibit the interaction of myostatin and GDF11 with the extracellular ligand-binding domain of their high affinity receptor, Activin receptor IIb. - in luciferase reporter assays both WFIKKN proteins are potent inhibitors of the signaling activity of myostatin and GDF11. - SPR interaction measurements with several additional representatives of TGF? family revealed that both WFIKKN1 and WFIKKN2 bind TGF?1, BMP2 and BMP4 with relatively high affinity (Kd ~ 10-7- 10-8 M). WFIKKN proteins showed weak affinity for growth factors BMP3 and BMP8b, and they did not bind to Activin A. - Despite their significant affinity for TGF?1, BMP2 and BMP4, in reporter assays neither WFIKKN1 nor WFIKKN2 inhibited the signaling activity of TGF?1, BMP2 and BMP4. In summary: our data indicate that WFIKKN proteins block the signalling activity of myostatin and GDF11 specifically and potently by sequestering these growth factors thereby preventing their binding to their receptors

Genome polymorphisms in HPV6s from benign respiratory and genital lesions

Complete genomes of HPV6s from respiratory papillomatoses and from a genital condyloma (single episode) were determined and compared to published genomes. Three adult onset respiratory papillomatoses (one solitary and two recurrent papillomas with two and six episodes) were HPV6b positive and five HPV6vcs (in the condyloma, in two juvenile papillomatoses with four and five recurrences and in two adult onset papillomatoses with seven and twelve episodes) were found, HPV6a was not encountered. In HPV6b, 25 polymorphisms were identified, 17 to 21 polymorphisms in a genome. Ten were virus-specific and fifteen were characteristic to the intratypic variant group. All three HPV6b genomes clustered separately from the prototype into three different groups. Five, two and two polymorphisms were found in E1, E5a and E6 ORFs, respectively, of which those of E5a were unique. These two resulted in amino acid alteration (E39D and P78S), others were silent. Other early ORFs were conservative. Late ORFs L1 and L2 contained four and five conservative polymorphisms, respectively. In the noncoding region one, in the long control region (LCR) six polymorphisms were detected. In HPV6vc, 22 polymorphisms were found, three to seven polymorphisms in a genome. Only one was present in all five genomes, one in three; 20 were unique. All five genomes clustered to the same large group as the reference genome, but all to different subclusters. ORFs E1, E2, E4, E5a and E6 carried three, five, one, one and two polymorphisms, respectively, of which, in contrast to HPV6b, all except two (in E1 and in E5a) were unique. Four of them led to amino acid replacement, all in the E2-E4 ORF (T116N, S144T, S246A and E340D in the E2; S246A corresponded to a S68R change in E4 ORF, the others were in the region belonging solely to E2). ORFs E5b and E7 were the same as the reference. Late ORF L1 contained one polymorphism common to all five genomes and five unique alterations. The common polymorphism led to a Y219D change. Three of the five unique polymorphisms were silent, one led to F441L and one to K449E amino acid replacement. In the noncoding region one unique, in the LCR two unique polymorphisms were detected. HPV6vc showed considerably higher variability with multiple non-silent polymorphisms in E2-E4, while coding regions of the three HPV6bs, though different from the prototype, were more similar. LCR, in contrast, was more variable in HPV6b. These suggest that HPV6bs differ mainly in LCR activity, while in HPV6vc polymorphisms of replication proteins may be more important. K. Szarka & G. Kardos were supported by a János Bolyai Research Scholarship of the Hungarian Academy of Science