{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T05:09:45Z","timestamp":1772168985466,"version":"3.50.1"},"update-to":[{"DOI":"10.12688\/f1000research.20135.1","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2020,3,16]],"date-time":"2020-03-16T00:00:00Z","timestamp":1584316800000}}],"reference-count":20,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2020,3,16]],"date-time":"2020-03-16T00:00:00Z","timestamp":1584316800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R21RR025412-01"],"award-info":[{"award-number":["R21RR025412-01"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Heinrich Pette Institute"},{"name":"Lovelace Respiratory Research Institute"}],"content-domain":{"domain":["f1000research.com"],"crossmark-restriction":false},"short-container-title":["F1000Res"],"abstract":"<ns4:p>\n                    <ns4:bold>Background:<\/ns4:bold>\n                    \u00a0The lack of adequate\n                    <ns4:italic>in vitro<\/ns4:italic>\n                    systems to isolate and propagate guinea pig adenovirus (GPAdV), a prevalent cause of respiratory illness of varaible severity in laboratory guinea pig colonies worldwide, has precluded its formal characterization to allow for the development of comprehensive diagnostic assays, and for the execution of complex pathogenesis and basic virology studies.\n                  <\/ns4:p>\n                  <ns4:p>\n                    <ns4:bold>Methods:<\/ns4:bold>\n                    Two strains of GPAdV were isolated in guinea pig (\n                    <ns4:italic>Cavia porcellus<\/ns4:italic>\n                    ) cell cultures from frozen archival infected animal tissue originated from colony outbreaks of pneumonia in Australia and the Czech Republic in 1996.\n                  <\/ns4:p>\n                  <ns4:p>\n                    <ns4:bold>Results:<\/ns4:bold>\n                    Commercially available guinea pig cell lines from colorectal carcinoma (GPC-16), fetal fibroblast (104-C1) and lung fibroblast (JH4 C1), and the tracheal epithelial cell line GPTEC-T developed in this study were able to support viral infection and early propagation. Sufficient viral DNA was recovered from cell cultures to PCR-amplify and obtain sequence data for the complete hexon gene and partial DNA polymerase and penton base genes. Phylogenetic analysis for the three regions of the genome provided strong evidence confirming GPAdV as a unique species in the genus Mastadenovirus.\n                  <\/ns4:p>\n                  <ns4:p>\n                    <ns4:bold>Conclusions:<\/ns4:bold>\n                    This study demonstrated the feasibility of propagating GPAdV in cultures of immortalized lines of GP cells of a variety of types, thus establishing a critical foundation for the development of a robust culture platform for virus stock production and titration. The generation and analysis of whole GPAdV genome sequences will provide additional data for a comprehensive description of the genetic organization of the viral genome and for a better assessment of genetic diversity between the two isolated strains.\n                  <\/ns4:p>","DOI":"10.12688\/f1000research.20135.2","type":"journal-article","created":{"date-parts":[[2020,3,16]],"date-time":"2020-03-16T07:30:06Z","timestamp":1584343806000},"page":"1597","update-policy":"https:\/\/doi.org\/10.12688\/f1000research.crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Isolation and initial propagation of guinea pig adenovirus (GPAdV) in Cavia porcellus cell lines"],"prefix":"10.12688","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6499-9579","authenticated-orcid":false,"given":"Adriana E.","family":"Kajon","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaoxin","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2180-2120","authenticated-orcid":false,"given":"Gabriel","family":"Gonzalez","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Susan","family":"Core","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Helga","family":"Hofmann-Sieber","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuguang","family":"Leng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"2560","published-online":{"date-parts":[[2020,3,16]]},"reference":[{"key":"ref-1","doi-asserted-by":"publisher","first-page":"488-91","DOI":"10.1177\/030098588302000411","article-title":"Naturally 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adenoviruses.","volume":"98","author":"B Tan","year":"2017","journal-title":"J Gen Virol."}],"container-title":["F1000Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/f1000research.com\/articles\/8-1597\/v2\/xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/f1000research.com\/articles\/8-1597\/v2\/pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/f1000research.com\/articles\/8-1597\/v2\/iparadigms","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,3,23]],"date-time":"2020-03-23T11:30:16Z","timestamp":1584963016000},"score":1,"resource":{"primary":{"URL":"https:\/\/f1000research.com\/articles\/8-1597\/v2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,3,16]]},"references-count":20,"URL":"https:\/\/doi.org\/10.12688\/f1000research.20135.2","relation":{"has-review":[{"id-type":"doi","id":"10.5256\/f1000research.25109.r61364","asserted-by":"subject"},{"id-type":"doi","id":"10.5256\/f1000research.25109.r61364","asserted-by":"object"}]},"ISSN":["2046-1402"],"issn-type":[{"value":"2046-1402","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,3,16]]},"assertion":[{"value":"Indexed","URL":"https:\/\/f1000research.com\/articles\/8-1597\/v2#article-reports","order":0,"name":"referee-status","label":"Referee status","group":{"name":"current-referee-status","label":"Current Referee Status"}},{"value":"10.5256\/f1000research.22110.r53516, M\u00e1rton Z. Vidovszky, Institute for Veterinary Medical Research, Centre for Agricultural Research, Budapest, Hungary, 18 Sep 2019, version 1, 1 approved, 1 approved with reservations","URL":"https:\/\/f1000research.com\/articles\/8-1597\/v1#referee-response-53516","order":0,"name":"referee-response-53516","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"<b>Adriana Kajon<\/b>; \n<i>Posted: 06 Mar 2020<\/i>; We would like to thank Dr. Vidovsky for his comments and suggestions for improvement of our paper. Below are our responses to each individual comment\/suggestion: &nbsp; This manuscript describes the isolation of two strains of guinea pig adenoviruses and their successful propagation on a newly developed and on three commercially available guinea pig (GP) cell lines. The manuscript is very relevant, as every successful, novel virus isolation is very important for adenovirology. The manuscript also discusses the partial phylogenetic comparison of adenoviral sequences with the examined AdV strains (full hexon gene and partial polymerase and penton base sequence parts). The Introduction is thorough enough and dense in a good meaning. It only deals with guinea pig AdVs, but all the known publications are well collected. -The Methods section is a bit too detailed and long compared to the other parts of the manuscript. Sometimes the process is already published somewhere, and the manuscript refers to it correctly but also details shortly. This is unnecessary in some cases in my opinion. Response: \n<i>The detailed description of methods and reagents is an F1000 Research requirement.<\/i> &nbsp; -The process of the primary cell line production and its immortalization is very well-written. It is easy to follow the process and very interesting. The successful isolation and propagation of the two GPAdVs are a very nice job and well written. The pictures of CPEs on different cell lines represent the changes correctly and well visualize this part of the research. The surprising parts of the adenoviral PCR detection in the homogenates and infected cell cultures are that the original nasal scraping sample was PCR negative for AdV, while the cell culture infected by this material showed CPE and it was PCR positive. I would be curious of the opinion of the authors, how this can be possible, i.e., that the PCR reaction is not enough sensitive for detection, but the material is suitable for the usually less successful virus isolation. Response: \n<i>We actually interpreted this result as a strong indicator that the original nasal scrapings sample contained infectious virus and that cell cultures were permissive for viral replication and amplification of the original input virus.<\/i> &nbsp; -It is a great result that the virus isolation attempts were successful with the GPAdV positive materials. It is more pity that the very interesting full genome sequence of these strains are not available. In the case of isolated viruses, it is only a small step by NGS. Because of the lack of full genome sequences of the examined AdVs, Figure 5 loses its relevancy. This makes some data incomparable in the figure. The phylogenetic calculations are very interesting, however, the one, based on the full hexon gene, seems to show a bit different topology than the other two. It is surprising, that among mastadenoviruses, not the rodent AdVs are the most ancient ones as usual, and they are not monophyletic with the GPAdVs. Summarizing, this is a well-written manuscript with very relevant and precious results. Response: \n<i>Since the submission of this manuscript, we have been working on obtaining whole genome sequences for GPAdV and expect to generate publications in the near future reporting those results.<\/i> &nbsp; Minor notes: -There are a couple of not disturbing misspellings or inadvertent mistakes. The only one, which can be confusing is in the legend of Figure 5. The labeling is correct on the figure, but in the legend, both columns are labeled with blue instead of orange and blue. Response: \n<i>We have corrected all spelling mistakes\/typos in the body of the manuscript and figure legends. <\/i> &nbsp; Other mistakes: -Misspelling in the abstract\/background: \u201cvariable\u201d instead of \u201cvariable\u201d. Twice, the word \u201cresuspended\u201d was misspelled to \u201cresuspened\u201d (Methods\/Immortalization, 1 paragraph). Also, here in the last sentence the word \u201caliquoted\u201d was misspelled to \u201calliquoted\u201d. In most of the times, it was used correctly, but several times the \u201cu\u201d character was used instead of\u201c&micro;\u201d e.g.: twice in Methods\/Immortalization, 2&nbsp; paragraph. Also, here a space is missing in one of the cases, between the number and the unit (1ug), correctly (1 &micro;g). The lack of space mistake between the amount and the unit occurs several times in the manuscript. I won't detail all of them here. Please check them. Also, a dot is unnecessary in the middle of a sentence of this paragraph, after a catalog nr: (catalog # MF10096). Response: \n<i>All of these corrections were incorporated into the revised version of the manuscript.<\/i> &nbsp; -A comma should be used in the listing instead of \u201cand\u201d at Methods\/GPTEC-T cell line authentication, after (DSMZ ACC 057). Response: \n<i>The comma was NOT inserted because cell line 104 C1 was NOT obtained from the DSMZ collection. The text was edited to read <b>\u201c... HeLa (DSMZ ACC 057) and 293 (DSMZ ACC 305), and cell line 104C1 (ATCC CRL-1405) were used as negative controls\u201d<\/b><\/i> &nbsp; -The shortening of fowl adenovirus D was used incorrectly. The letter \u201ca\u201d of adenovirus is in lower case font. It should be a capital \u201cA\u201d. Everywhere else it was used correctly. (Methods\/Assembly, 1 paragraph: \u201cFadV-D\u201d was written instead of \u201cFAdV-D\u201d). Once, the word \u201cadenovirus\u201d was written also with a capital incorrectly in the case of cynomolgus adenovirus. A \u201c3\u2019\u201d end is missing for primer GPhexFW3 in Table 1. The word \u201ccytopathic\u201d was misspelled to \u201ccythopathyc\u201d (legend of Figure 3.). The word \u201ctwenty\u201d was misspelled to \u201ctwety\u201d (legend of Figure 4.). Also, here the word \u201cabsence\u201d was misspelled to \u201cabsense\u201d. The use of abbreviation AdV species with the number of type like HAdV-C5 is not exactly appropriate, but widespread, so it is okay. But once it was misspelled (Methods\/Assembly, 2 paragraph: \u201cHAdVA-12\u201d instead of \u201cHAdV-A12\u201d). Also here in the whole paragraph, while in abbreviation the first letter is indeed in capital like \u201cHAdVA\u201d, in writing the names of viruses should be written by lower case like \u201chuman adenovirus A12\u201d according to the recommendation of the ICTV (if the name is from a Latin genus\/species name, like Odocoileus hemionus adenovirus, it is correct with capital). The name of fowl adenovirus in Methods\/Assembly and in the legend of Figure 4 is incorrect (species D Fowl aviadenovirus 764). The correct name should be something like: \u201cfowl adenovirus 8b strain 764 (FAdV-D)\u201d. Response: \n<i>All of these corrections were incorporated into the revised version of the manuscript.<\/i> &nbsp; -I would prefer the use of AdV type names (Arabic numbers) instead of species names (letters) on the phylogenetic trees, and GenBank numbers instead of RefSeq identification numbers (like starting with NC_...). These latter data won\u2019t be updated, while the GenBank entries may be. Based on this, type data (including AC_... data) are more appropriate here. Response: \n<i>The original designations NC_034834 and NC_000942, were changed to KY306667 and AC_000012, respectively as requested in the Methods section and in revised Figures 4 and 5.<\/i> &nbsp; -The examined AdVs are originated from rodent hosts. I missed very much other available AdVs with rodent origin on the phylogenetic trees, like squirrel AdV (KY427939), or murine adenovirus 2 (HM049560). If the data was too much for the calculation, I would skip rather MAdV-1 or MAdV-3. They are more similar to each other, while MAdV-2 is more different. Response: \n<i>We completely agree with Dr. Vidovszky\u2019s comments. We have made recent observations of close similarities between the genomes of GPAdV and MAdV-2 so the revisions addressing his comments are extremely important and timely. We have therefore incorporated the sequences for MAdV-2 into our new analyses for figures 4 and 5 and have revised both figures, and underlying data file now designated \u201c<\/i>GC Content values 030320.xls\u201d\n<i>. <\/i>","URL":"https:\/\/f1000research.com\/articles\/8-1597\/v2#referee-comment-5283","order":1,"name":"referee-comment-5283","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.25109.r61364, Ana Paula Ravazzolo, Faculdade de Veterin\u00e1ria, Laborat\u00f3rio de Imunologia e Biologia Molecular, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, Brazil, 23 Mar 2020, version 2, indexed","URL":"https:\/\/f1000research.com\/articles\/8-1597\/v2#referee-response-61364","order":2,"name":"referee-response-61364","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"This work was partially supported by the US National Institute of Health (NIH) grant R21 RR025412-01 to author AEK, and by intramural funding from the Lovelace Respiratory Research Institute and the Heinrich Pette Institute, Leibniz Institute for Experimental Virology. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.","order":3,"name":"grant-information","label":"Grant Information"},{"value":"This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.","order":0,"name":"copyright-info","label":"Copyright"}]}}