Lactococcus lactis expressing sand fly PpSP15 salivary protein confers long-term protection against Leishmania major in BALB/c mice
Cutaneous leishmaniasisis a vector-borne disease transmitted by Leishmania infected sand flies. PpSP15 is an immunogenic salivary protein from the sand fly Phlebotomus papatasi. Immunization with PpSP15 was shown to protect against Leishmania major infection. Lactococcus lactis is a safe non-pathoge...
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| creator | Davarpanah, Elaheh Seyed, Negar Bahrami, Fariborz Rafati, Sima Safaralizadeh, Reza Taheri, Tahereh |
| description | Cutaneous leishmaniasisis a vector-borne disease transmitted by Leishmania infected sand flies. PpSP15 is an immunogenic salivary protein from the sand fly Phlebotomus papatasi. Immunization with PpSP15 was shown to protect against Leishmania major infection. Lactococcus lactis is a safe non-pathogenic delivery system that can be used to express antigens in situ. Here, the codon-optimized Ppsp15-egfp gene was cloned in pNZ8121 vector downstream of the PrtP signal peptide that is responsible for expression and secretion of the protein on the cell wall. Expression of PpSP15-EGFP recombinant protein was monitored by immunofluorescence, flow cytometry and Western blot. Also, expression of protein in cell wall compartment was verified using whole cell ELISA, Western blot and TEM microscopy. BALB/c mice were immunized three times with recombinant L. lactis-PpSP15-EGFPcwa, and the immune responses were followed up, at short-term (ST, 2 weeks) and long-term (LT, 6 months) periods. BALB/c mice were challenged with L. major plus P. papatasi Salivary Gland Homogenate. Evaluation of footpad thickness and parasite burden showed a delay in the development of the disease and significantly decreased parasite numbers in PpSP15 vaccinated animals as compared to control group. In addition, immunized mice showed Th1 type immune responses. Importantly, immunization with L. lactis-PpSP15-EGFPcwa stimulated the long-term memory in mice which lasted for at least 6 months. |
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PpSP15 is an immunogenic salivary protein from the sand fly Phlebotomus papatasi. Immunization with PpSP15 was shown to protect against Leishmania major infection. Lactococcus lactis is a safe non-pathogenic delivery system that can be used to express antigens in situ. Here, the codon-optimized Ppsp15-egfp gene was cloned in pNZ8121 vector downstream of the PrtP signal peptide that is responsible for expression and secretion of the protein on the cell wall. Expression of PpSP15-EGFP recombinant protein was monitored by immunofluorescence, flow cytometry and Western blot. Also, expression of protein in cell wall compartment was verified using whole cell ELISA, Western blot and TEM microscopy. BALB/c mice were immunized three times with recombinant L. lactis-PpSP15-EGFPcwa, and the immune responses were followed up, at short-term (ST, 2 weeks) and long-term (LT, 6 months) periods. BALB/c mice were challenged with L. major plus P. papatasi Salivary Gland Homogenate. Evaluation of footpad thickness and parasite burden showed a delay in the development of the disease and significantly decreased parasite numbers in PpSP15 vaccinated animals as compared to control group. In addition, immunized mice showed Th1 type immune responses. Importantly, immunization with L. lactis-PpSP15-EGFPcwa stimulated the long-term memory in mice which lasted for at least 6 months.</description><identifier>ISSN: 1935-2735</identifier><identifier>ISSN: 1935-2727</identifier><identifier>EISSN: 1935-2735</identifier><identifier>DOI: 10.1371/journal.pntd.0007939</identifier><identifier>PMID: 31899767</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Animals ; Antigens ; Bacteria ; Biology and Life Sciences ; Cell walls ; Design ; Disease transmission ; Drug resistance ; ELISA ; Enzyme-linked immunosorbent assay ; Female ; Flow cytometry ; Genes ; Immune response ; Immunization ; Immunofluorescence ; Immunogenicity ; Immunological memory ; Immunotherapy ; Infectious diseases ; Insect Proteins - immunology ; Lactococcus lactis ; Lactococcus lactis - genetics ; Lactococcus lactis - metabolism ; Leishmania major ; Leishmaniasis, Cutaneous - transmission ; Long term memory ; Lymphocytes T ; Medical research ; Medicine and Health Sciences ; Mice, Inbred BALB C ; Microscopy ; Parasites ; Parasitic diseases ; Pathogens ; Phlebotomus - genetics ; Proteins ; Recombinant Proteins - immunology ; Recombinants ; Research and Analysis Methods ; Salivary gland ; Salivary glands ; Salivary Proteins and Peptides - genetics ; Salivary Proteins and Peptides - metabolism ; Secretion ; Thickness ; Tropical diseases ; Vaccines ; Vector-borne diseases</subject><ispartof>PLoS neglected tropical diseases, 2020-01, Vol.14 (1), p.e0007939-e0007939</ispartof><rights>2020 Davarpanah et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2020 Davarpanah et al 2020 Davarpanah et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c526t-f0292e01d8b73369a9bd880b23d7c92dcb32f0e5152f2719c732a53b284c1c8a3</citedby><cites>FETCH-LOGICAL-c526t-f0292e01d8b73369a9bd880b23d7c92dcb32f0e5152f2719c732a53b284c1c8a3</cites><orcidid>0000-0002-2619-6879</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6941807/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6941807/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2100,2926,23864,27922,27923,53789,53791,79370,79371</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31899767$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Bueno, Lilian Lacerda</contributor><creatorcontrib>Davarpanah, Elaheh</creatorcontrib><creatorcontrib>Seyed, Negar</creatorcontrib><creatorcontrib>Bahrami, Fariborz</creatorcontrib><creatorcontrib>Rafati, Sima</creatorcontrib><creatorcontrib>Safaralizadeh, Reza</creatorcontrib><creatorcontrib>Taheri, Tahereh</creatorcontrib><title>Lactococcus lactis expressing sand fly PpSP15 salivary protein confers long-term protection against Leishmania major in BALB/c mice</title><title>PLoS neglected tropical diseases</title><addtitle>PLoS Negl Trop Dis</addtitle><description>Cutaneous leishmaniasisis a vector-borne disease transmitted by Leishmania infected sand flies. PpSP15 is an immunogenic salivary protein from the sand fly Phlebotomus papatasi. Immunization with PpSP15 was shown to protect against Leishmania major infection. Lactococcus lactis is a safe non-pathogenic delivery system that can be used to express antigens in situ. Here, the codon-optimized Ppsp15-egfp gene was cloned in pNZ8121 vector downstream of the PrtP signal peptide that is responsible for expression and secretion of the protein on the cell wall. Expression of PpSP15-EGFP recombinant protein was monitored by immunofluorescence, flow cytometry and Western blot. Also, expression of protein in cell wall compartment was verified using whole cell ELISA, Western blot and TEM microscopy. BALB/c mice were immunized three times with recombinant L. lactis-PpSP15-EGFPcwa, and the immune responses were followed up, at short-term (ST, 2 weeks) and long-term (LT, 6 months) periods. BALB/c mice were challenged with L. major plus P. papatasi Salivary Gland Homogenate. Evaluation of footpad thickness and parasite burden showed a delay in the development of the disease and significantly decreased parasite numbers in PpSP15 vaccinated animals as compared to control group. In addition, immunized mice showed Th1 type immune responses. 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PLoS neglected tropical diseases</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Davarpanah, Elaheh</au><au>Seyed, Negar</au><au>Bahrami, Fariborz</au><au>Rafati, Sima</au><au>Safaralizadeh, Reza</au><au>Taheri, Tahereh</au><au>Bueno, Lilian Lacerda</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Lactococcus lactis expressing sand fly PpSP15 salivary protein confers long-term protection against Leishmania major in BALB/c mice</atitle><jtitle>PLoS neglected tropical diseases</jtitle><addtitle>PLoS Negl Trop Dis</addtitle><date>2020-01-01</date><risdate>2020</risdate><volume>14</volume><issue>1</issue><spage>e0007939</spage><epage>e0007939</epage><pages>e0007939-e0007939</pages><issn>1935-2735</issn><issn>1935-2727</issn><eissn>1935-2735</eissn><abstract>Cutaneous leishmaniasisis a vector-borne disease transmitted by Leishmania infected sand flies. PpSP15 is an immunogenic salivary protein from the sand fly Phlebotomus papatasi. Immunization with PpSP15 was shown to protect against Leishmania major infection. Lactococcus lactis is a safe non-pathogenic delivery system that can be used to express antigens in situ. Here, the codon-optimized Ppsp15-egfp gene was cloned in pNZ8121 vector downstream of the PrtP signal peptide that is responsible for expression and secretion of the protein on the cell wall. Expression of PpSP15-EGFP recombinant protein was monitored by immunofluorescence, flow cytometry and Western blot. Also, expression of protein in cell wall compartment was verified using whole cell ELISA, Western blot and TEM microscopy. BALB/c mice were immunized three times with recombinant L. lactis-PpSP15-EGFPcwa, and the immune responses were followed up, at short-term (ST, 2 weeks) and long-term (LT, 6 months) periods. BALB/c mice were challenged with L. major plus P. papatasi Salivary Gland Homogenate. Evaluation of footpad thickness and parasite burden showed a delay in the development of the disease and significantly decreased parasite numbers in PpSP15 vaccinated animals as compared to control group. In addition, immunized mice showed Th1 type immune responses. Importantly, immunization with L. lactis-PpSP15-EGFPcwa stimulated the long-term memory in mice which lasted for at least 6 months.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>31899767</pmid><doi>10.1371/journal.pntd.0007939</doi><orcidid>https://orcid.org/0000-0002-2619-6879</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Antigens Bacteria Biology and Life Sciences Cell walls Design Disease transmission Drug resistance ELISA Enzyme-linked immunosorbent assay Female Flow cytometry Genes Immune response Immunization Immunofluorescence Immunogenicity Immunological memory Immunotherapy Infectious diseases Insect Proteins - immunology Lactococcus lactis Lactococcus lactis - genetics Lactococcus lactis - metabolism Leishmania major Leishmaniasis, Cutaneous - transmission Long term memory Lymphocytes T Medical research Medicine and Health Sciences Mice, Inbred BALB C Microscopy Parasites Parasitic diseases Pathogens Phlebotomus - genetics Proteins Recombinant Proteins - immunology Recombinants Research and Analysis Methods Salivary gland Salivary glands Salivary Proteins and Peptides - genetics Salivary Proteins and Peptides - metabolism Secretion Thickness Tropical diseases Vaccines Vector-borne diseases |
| title | Lactococcus lactis expressing sand fly PpSP15 salivary protein confers long-term protection against Leishmania major in BALB/c mice |
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