Federico Zuckermann - Profile on Academia.edu (original) (raw)

Papers by Federico Zuckermann

Bacillus-based direct-fed microbial reduces the pathogenic synergy of a co-infection with Salmonella enterica serovar Choleraesuis and porcine reproductive and respiratory syndrome virus

Raw data and its analysis collected from a trial designed to test the impact of providing a Bacil... more Raw data and its analysis collected from a trial designed to test the impact of providing a Bacillus-based direct-fed microbial (DFM) on the syndrome resulting from orally infecting pigs with either Salmonella enterica serotype Choleraesuis (S. Choleraesuis) alone, or in combination with an intranasal challenge, three days later, with porcine reproductive and respiratory syndrome virus (PRRSV).

Functional and phenotypic analysis of porcine peripheral blood CD4/CD8 double-positive T cells

PubMed, Mar 1, 1996

Functional and phenotypic properties of porcine peripheral blood CD4/CD8 double-positive (DP) lym... more Functional and phenotypic properties of porcine peripheral blood CD4/CD8 double-positive (DP) lymphocytes were examined. In cross-sectional and longitudinal studies involving a total of 103 pigs, this lymphocyte population was found to increase gradually in proportion with age, comprising < 2% of the total peripheral blood lymphocyte pool in 1-week-old swine and reaching 30-55% by 3 years of age. CD4/CD8 DP lymphocytes were able to proliferate in response to stimulation with recall viral antigen. Furthermore, these cells mostly expressed high levels of the surface antigen recognized by monoclonal antibody (mAb) 4B4 (4B4hi), which is specific for the human beta 1 integrin. The CD4+4B4hi lymphocytes from pseudorabies virus-immune swine, proliferated in response to stimulation with the homologous virus, while CD4+4B4lo lymphocytes did not. Stimulation of CD4 single-positive (SP) cells with recall viral antigen, but not with mitogen, resulted in the generation of lymphoblasts which were predominantly of CD4/CD8 DP phenotype, suggesting a role for recall antigen in the generation of this lymphocyte subset. More than half of the CD4+ lymphocytes from palatine tonsils of 6-month-old swine were CD4/CD8 DP, while in the lymph nodes CD4/CD8 DP cells accounted for only one-third or less of CD4+ cells. In contrast, CD4/CD8 DP lymphocytes were absent from the palatine tonsils of 3-day-old swine, which only contained CD4 SP cells. Together, these results indicate that porcine CD4/CD8 DP lymphocytes, exhibit properties of mature antigen-experienced cells, and are inducible by stimulation with recall antigen. These data are consistent with the hypothesis that this population in swine includes memory/effector T cells.

Journal of Virology, Feb 1, 1990

Pseudorabies virus (PrV) is the etiological agent of Aujeszky's disease, a disease that causes he... more Pseudorabies virus (PrV) is the etiological agent of Aujeszky's disease, a disease that causes heavy economic losses in the swine industry. A rational approach to the generation of an effective vaccine against this virus requires an understanding of the immune response induced by it and of the role of the various viral antigens in inducing such a response. We have constructed mutants of PrV [strain PrV(Ka)] that differ from each other only in expression of the viral nonessential glycoproteins gI, gp63, gX, and glll (i.e., are otherwise isogenic). These mutants were used to ascertain the importance of each of the nonessential glycoproteins in eliciting a PrV-specific cytotoxic T-lymphocyte (CTL) response in mice and pigs. Immunization of DBA/2 mice and pigs with a thymidine kinase-deficient (TK-) mutant of PrV elicits the formation of cytotoxic cells that specifically lyse syngeneic infected target cells. These PrV-specific cytolytic cells have the phenotype of major histocom- patibility complex class I antigen-restricted CTLs. The relative number of CTLs specific for glycoproteins gI, gp63, gX, and gIll induced in mice vaccinated with a TK-mutant of PrV was ascertained by comparing their levels of cytotoxicity against syngeneic cells infected with either wild-type virus or gI-/gp63-, gX-, or gi- virus deletion mutants. The PrV-specific CLTs were significantly less effective in lysing gi--infected targets than in lysing gI-/gp63-, gX-, or wild-type-infected targets. The in vitro secondary CTL response of lymphocytes obtained from either mice or pigs 6 or more weeks after immunization with a TK-mutant of PrV was also tested. Lymphocytes obtained from these animals were cultured with different glycoprotein-deficient mutants of PrV, and their cytolytic activities against wild-type-infected targets were ascertained. The importance of each of the nonessential viral glycoproteins in eliciting CTLs was assessed from the effectiveness of each of the virus mutants to stimulate the secondary anti-PrV CTL response. Cultures of both murine or swine lymphocytes that had been stimulated with gIIIvirus contained only approximately half as many lytic units as did those stimulated with either wild-type virus, a gXvirus mutant, or a gl-/gp63virus mutant. Thus, a large proportion of the PrV-specific CTLs that are induced by immunization with PrV of both mice and pigs are directed against gIll. Furthermore, glycoproteins gI, gp63, and gX play at most a minor role in the CTL response of these animals to PrV.

Journal of Virology, Jan 15, 2018

Porcine reproductive and respiratory syndrome virus (PRRSV) infects alveolar macrophages (AM), ca... more Porcine reproductive and respiratory syndrome virus (PRRSV) infects alveolar macrophages (AM), causing dysregulated alpha interferon (IFN-␣) and tumor necrosis factor alpha (TNF-␣) production through a mechanism(s) yet to be resolved. Here, we show that AM infected with PRRSV secreted a reduced quantity of IFN-␣ following exposure of the cell to synthetic double-stranded RNA (dsRNA). This reduction did not correlate with reduced IFNA1 gene transcription. Rather, it coincided with two events that occurred late during infection and that were indicative of translational attenuation, specifically, the activation of eukaryotic translation initiation factor 2␣ (eIF2␣) and the appearance of stress granules. Notably, the typical rapid production of TNF-␣ by AM exposed to lipopolysaccharide (LPS) was suppressed or enhanced by PRRSV, depending on when the LPS exposure occurred after virus infection. If exposure was delayed until 6 h postinfection (hpi) so that the development of the cytokine response coincided with the time in which phosphorylation of eIF2␣ by the stress sensor PERK (protein kinase RNA [PKR]-like ER kinase) occurred, inhibition of TNF-␣ production was observed. However, if LPS exposure occurred at 2 hpi, prior to a detectable onset of eIF2␣ phosphorylation, a synergistic response was observed due to the earlier NF-B activation via the stress sensor IRE1␣ (inositol-requiring kinase 1␣). These results suggest that the asynchronous actions of two branches of the unfolded protein response (UPR), namely, IRE1␣, and PERK, activated by ER stress resulting from the virus infection, are associated with enhancement or suppression of TNF-␣ production, respectively. IMPORTANCE The activation of AM is controlled by the microenvironment to deter excessive proinflammatory cytokine responses to microbes that could impair lung function. However, viral pneumonias frequently become complicated by secondary bacterial infections, triggering severe inflammation, lung dysfunction, and death. Although dysregulated cytokine production is considered an integral component of the exacerbated inflammatory response in viral-bacterial coinfections, the mechanism responsible for this event is unknown. Here, we show that PRRSV replication in porcine AM triggers activation of the IRE1␣ branch of the UPR, which causes a synergistic TNF-␣ response to LPS exposure. Thus, the severe pneumonias typically observed in pigs afflicted with PRRSV-bacterial coinfections could result from dysregulated, overly robust TNF-␣ production in response to opportunistic pathogens that is not commensurate with the typical restrained reaction by uninfected AM. This notion could help in the design of therapies to mitigate the severity of viral and bacterial coinfections.

Journal of Immunology, Jun 1, 1999

We previously identified four novel cDNA fragments related to human esophageal cancer. One of the... more We previously identified four novel cDNA fragments related to human esophageal cancer. One of the fragments was named esophageal cancer related gene 2 (ECRG2). We report here the molecular cloning, sequencing, and expression of the ECRG2 gene. The ECRG2 cDNA comprises a 258 bp nucleotide sequence which encodes for 85 amino acids with a predicted molecular weight of 9.2 kDa. Analysis of the protein sequence reveals the presence at the N terminus of a signal peptide followed by 56 amino acids with a significant degree of sequence similarity with the conserved Kazal domain which characterizes the serine protease inhibitor family. Pulse-chase experiments showed that ECRG2 protein was detected in both cell lysates and culture medium, indicating that the ECRG2 protein was extracellularly secreted after the post-translational cleavage. In vitro uPA/plasmin activity analysis showed the secreted ECRG2 protein inhibited the uPA/plasmin activity, indicating that ECRG2 may be a novel serine protease inhibitor. Northern blot analysis revealed the presence of the major band corresponding to a size of 569 kb throughout the fetal skin, thymus, esophagus, brain, lung, heart, stomach, liver, spleen, colon, kidney, testis, muscle, cholecyst tissues and adult esophageal mucosa, brain, thyroid tissue and mouth epithelia. However, ECRG2 gene was significantly downregulated in primary esophageal cancer tissues. Taken together, these results indicate that ECRG2 is a novel member of the Kazal-type serine protease inhibitor family and may function as a tumor suppressor gene regulating the protease cascades during carcinogenesis and migration/invasion of esophageal cancer.

Analyses of monoclonal antibodies reactive with porcine CD44 and CD45

Veterinary Immunology and Immunopathology, Oct 1, 1994

... Binns b , Robert Husmann a , Huaizhi Yang c , Margaret M. Carr d , Yoon Berm Kim e , William ... more ... Binns b , Robert Husmann a , Huaizhi Yang c , Margaret M. Carr d , Yoon Berm Kim e , William ... been observed, of molecular weights 190, 210, 225 kDa and 180, 205, 220 kDa, re-spectively (Maddox ... The mAb W# 122 (LIL 13 ), had a reac-tivity and staining pattern similar to the ...

Expression of MHC antigens on murine trophoblast and their modulation by interferon

Journal of Immunology, Aug 1, 1986

Primary cultures of defined populations of mouse trophoblast, isolated from mature placentas, wer... more Primary cultures of defined populations of mouse trophoblast, isolated from mature placentas, were analyzed for their MHC antigen expression and for the modulatory effect of interferon (IFN) by antibody- and complement-mediated cytotoxicity and flow cytofluorometry. The cells were obtained from placentas by enzymatic digestion, followed by Percoll gradient fractionation, and are large, fetally derived epithelial cells, which we previously characterized and identified as trophoblast cells. After 2 days in culture, a significant proportion of the trophoblast cells were susceptible to antibody- and complement-mediated lysis by anti-paternal strain alloantisera (40%) and, to a lesser degree, by an anti-class I monoclonal antibody (20%). Flow cytofluorometric analysis indicated that 20 to 50% of the cultured trophoblast cells expressed low levels of paternal strain class I antigens as compared to L cell fibroblasts. After culture for 48 hr with IFN-alpha/beta or IFN-gamma, the percent of class I-positive cells was increased to 68 to 76%, as was the mean fluorescence intensity, which correlated with the increased percent of antibody- and complement-mediated specific lysis (73%). No expression of class II MHC antigen by the cultured trophoblast cells was detected, even after culture in the presence of IFN-gamma. The cultured trophoblast cells, when tested for alkaline phosphatase (AP) activity, were composed of strongly positive and weakly positive subpopulations. An inverse correlation between strength of AP activity and the expression of H-2 was observed by double staining. These results indicate that trophoblast cells cultured in vitro are able to express paternal strain class I but not class II MHC antigens, as has been reported in vivo, and that this expression can be modulated by IFN. Further study of these cells should provide important clues for the understanding of materno-fetal coexistence in the face of MHC antigen differences.

Emerging microbes & infections, 2020

The main target cells for African swine fever virus (ASFV) replication in pigs are of monocyte ma... more The main target cells for African swine fever virus (ASFV) replication in pigs are of monocyte macrophage lineage and express markers typical of the intermediate to late stages of differentiation. The lack of a porcine cell line, which accurately represents these target cells, limits research on virus host interactions and the development of liveattenuated vaccine strains. We show here that the continuously growing, growth factor dependent ZMAC-4 porcine macrophage cell line is susceptible to infection with eight different field isolates of ASFV. Replication in ZMAC-4 cells occurred with similar kinetics and to similar high titres as in primary porcine bone marrow cells. In addition we showed that twelve passages of an attenuated strain of ASFV, OURT88/3, in ZMAC-4 cells did not reduce the ability of this virus to induce protection against challenge with virulent virus. Thus, the ZMAC-4 cells provide an alternative to primary cells for ASFV replication.

Veterinary Sciences, Mar 13, 2023

Rapid diagnostics for viral disease in livestock are urgently needed, especially for those that p... more Rapid diagnostics for viral disease in livestock are urgently needed, especially for those that pose threats to human health and/or food security. Influenza A virus is a zoonotic disease that has a large reservoir in commercial swine herds. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a tool that has the potential to detect influenza A virus in easy-to-obtain swine nasal samples in a short time period, with minimal laboratory equipment. This article outlines how to process nasal samples and complete an RT-LAMP assay in a laboratory setting, demonstrating a protocol that goes from sample collection to positive test results in less than one hour. Ultimately, this diagnostic test system can be developed for producers to identify sick animals on-farm, and to avoid logistical issues and time delays associated with the use of diagnostic laboratories. In addition, rapid results allow producers to implement interventions in real time, thus enhancing the efficacy of preventative measures and mitigating outbreaks.

Journal of Immunology, Nov 1, 1987

Journal of Virology, Dec 1, 1988

To ascertain the biological functions of different glycoproteins that are nonessential for pseudo... more To ascertain the biological functions of different glycoproteins that are nonessential for pseudorabies virus growth in vitro, we have constructed mutants defective in one (or a combination) of these glycoproteins and have examined various aspects of their role in the infective process. We made the following two observations. (i) Glycoproteins gI and gp63 are noncovalently complexed to each other. They are coprecipitated by antisera against either one of these glycoproteins but do not share antigenic determinants: monoclonal antibodies against gp63 do not immunoprecipitate gI from extracts of gp63mutant-infected cells, and monoclonal antibodies against gI do not immunoprecipitate gp63 from extracts of glmutant-infected cells. (ii) Mutants unable to synthesize either gI or gp63 have some common biological characteristics; they have a growth advantage in primary chicken embryo fibroblasts. Furthermore, we have shown previously that in conjunction with glycoprotein gIII, gI and gp63 are necessary for the expression of virulence (T. C. Mettenleiter, C.

PRRSV: comparación de vacunos comerciales respecto a su capacidad para inducir protección frente a cepas actuales de PRRSV de alta virulencia

Anaporc: revista de la Asociación de Porcinocultura Científica, 2004

Archives of Virology, Mar 23, 2016

Porcine reproductive and respiratory syndrome (PRRS) is a leading cause of economic burden to the... more Porcine reproductive and respiratory syndrome (PRRS) is a leading cause of economic burden to the pork industry worldwide. The routinely used modified live PRRS virus vaccine (PRRS-MLV) induces clinical protection, but it has safety concerns. Therefore, in an attempt to develop a safe and protective inactivated PRRSV vaccine, we generated PRRS-virus-like-particles (PRRS-VLPs) containing the viral surface proteins GP5-GP4-GP3-GP2a-M or GP5-M using a novel baculovirus expression system. Our in vitro results indicated that the desired PRRSV proteins were incorporated in both the VLPs preparations based on their reactivity in immunogold electron microscopy and ELISA. To boost their immunogenicity in pigs, we entrapped the PRRS-VLPs in PLGA nanoparticles and coadministered them intranasally with a potent adjuvant. We then evaluated their efficacy in pigs against a viral challenge using a virulent heterologous field isolate. Our results indicated that PRRS-VLPs induced an anamnestic immune response, since we observed boosted IgG and IFN-c production in vaccinated and virus-challenged animals, but not during the prechallenge period. Importantly, a two-log reduction in the lung viral load was detected in PRRS-VLP-vaccinated animals. In conclusion, we generated PRRS-VLPs containing up to five viral surface proteins and demonstrated their immunogenicity in pigs, but further studies are required to improve its immunogenicity and efficacy as a vaccine candidate.

Effect of transportation and feed withdrawal on shedding of Salmonella typhimurium among experimentally infected pigs

PubMed, Sep 1, 1999

Objective: To determine whether stress associated with transportation or feed withdrawal increase... more Objective: To determine whether stress associated with transportation or feed withdrawal increased fecal shedding of Salmonella Typhimurium among pigs experimentally infected with the organism. Animals: 86 healthy pigs. Procedure: Pigs were challenge exposed with Salmonella Typhimurium at 4 weeks old and reared conventionally. When pigs reached market weight, they were assigned to groups and subjected to various combinations of transportation and feed withdrawal. Ileocecal contents were collected after slaughter and tested for Salmonella Typhimurium. Results: Salmonella Typhimurium was not detected in feces collected from pigs just prior to slaughter. When feed was withheld for 24 hours prior to slaughter, the proportion of transported pigs with Salmonella Typhimurium in ileocecal contents at the time of slaughter was not significantly different from the proportion of nontransported pigs. However, when feed was not withheld prior to slaughter, the proportion of transported pigs with Salmonella Typhimurium in ileocecal contents at the time of slaughter was significantly higher than the proportion of nontransported pigs. Conclusions and clinical relevance: When carrier pigs remained on feed, transportation stress increased the proportion positive for Salmonella sp. On the basis of results reported here, it is suggested that producers withhold feed from pigs for 24 hours prior to transportation to a slaughter plant.

Second international swine CD workshop summary

The FASEB Journal, 1996

Journal of Virology, 1988

Deletion mutants of pseudorabies virus unable to express glycoprotein gIII, gI, or gp63 or double... more Deletion mutants of pseudorabies virus unable to express glycoprotein gIII, gI, or gp63 or double and triple mutants defective in these glycoproteins were constructed, and their virulence for day-old chickens inoculated intracerebrally was determined. Mutants of wild-type pseudorabies virus defective in glycoprotein gIII, gI, or gp63 were only slightly less virulent (at most, fivefold) for chickens than was the wild-type virus. However, mutants defective in both gIII and gI or gIII and gp63 were avirulent for chickens, despite their ability to grow in cell culture in vitro to about the same extent as mutants defective in gIII alone (which were virulent). These results show that gIII plays a role in virulence and does so in conjunction with gI or gp63. The effect of gIII on virulence was also shown when the resident gIII gene of variants of the Bartha vaccine strain (which codes for gIIIB) was replaced with a gIII gene derived from a virulent wild-type strain (which codes for gIIIKa)...

Virus Research, 2020

This is a PDF file of an article that has undergone enhancements after acceptance, such as the ad... more This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Journal of Virology, 1992

Mutants of pseudorabies virus defective in either glycoprotein gI or gIII are only slightly less ... more Mutants of pseudorabies virus defective in either glycoprotein gI or gIII are only slightly less virulent for mice and chickens than is wild-type virus, while mutants defective in both gI and gIII are avirulent. To clarify the reason for the lack of virulence of the gI- gIII- mutants, we have analyzed in some detail the interactions of these mutants with their hosts. The results obtained showed that the gI glycoprotein is an accessory protein that promotes cell fusion. This conclusion is based on the findings that in some cell types, syncytium formation is significantly reduced in mutants deficient in gI. Furthermore, despite efficient replication, gI- mutants form significantly smaller plaques on some cell types. Finally, while wild-type and gI- virus are neutralized similarly by antisera, the size of the plaques formed by gI- mutants, but not by wild-type virus, is reduced by the presence of neutralizing antibodies in the overlay. Passive immunization of mice with neutralizing ant...

Journal of Virology, 1989

Adsorption of mutants of pseudorabies virus (PrV) lacking glycoprotein gIII is slower and less ef... more Adsorption of mutants of pseudorabies virus (PrV) lacking glycoprotein gIII is slower and less efficient than is that of wild-type virus (C. Schreurs, T. C. Mettenleiter, F. Zuckermann, N. Snugg, and T. Ben-Porat, J. Virol. 62:2251-2257, 1988). To ascertain the functions of gIII in the early interactions of PrV with its host cells, we compared the effect on wild-type virus and gIII- mutants of antibodies specific for various PrV proteins. Although adsorption of wild-type virus was inhibited by polyvalent antisera against PrV as well as by sera against gIII and gp50 (but not sera against gII), adsorption of the gIII- mutants was not inhibited by any of these antisera. These results suggest that, in contrast to adsorption of wild-type PrV, the initial interactions of the gIII- mutants with their host cells are not mediated by specific viral proteins. Furthermore, competition experiments showed that wild-type Prv and the gIII- mutants do not compete for attachment to the same cellular ...

Journal of Virology, 1987

The Bartha vaccine strain of pseudorabies virus has a deletion in the short unique (Us) region of... more The Bartha vaccine strain of pseudorabies virus has a deletion in the short unique (Us) region of its genome which includes the genes that code for glycoproteins gI and gp63 (E. Petrovskis, J. G. Timmins, T. M. Gierman, and L. E. Post, J. Virol. 60:1166-1169, 1986). Restoration of an intact Us to the Bartha strain enhances its ability to be released from infected rabbit kidney cells and increases the size of the plaques formed on these cells (T. Ben-Porat, J. M. DeMarchi, J. Pendrys, R. A. Veach, and A. S. Kaplan, J. Virol. 57:191-196, 1986). To determine which gene function plays a role in virus release from rabbit kidney cells, deletions were introduced into the genomes of both wild-type virus and the "rescued" Bartha strain (Bartha strain to which an intact Us had been restored) that abolish the expression of either the gI gene alone or both gI and gp63 genes. The effect of these deletions on the phenotype of the viruses was studied. Deletion mutants of wild-type virus ...