Enhanced Surfactant Protein and Defensin mRNA Levels and Reduced Viral Replication during Parainfluenza Virus Type 3 Pneumonia in Neonatal Lambs

Grubor, Branka; Gallup, Jack M.; Meyerholz, David K.; Crouch, Erika C.; Evans, Richard B.; Brogden, Kim A.; Lehmkuhl, Howard D.; Ackermann, Mark R.

doi:10.1128/cdli.11.3.599-607.2004

Cited by 77 publications

(93 citation statements)

References 49 publications

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“…The lack of observable increase in lung SBD-1 mRNA levels during experimental pneumonia caused by M. haemolytica is contrary to our findings that demonstrate increased SBD-1 and surfactant protein A and D mRNA expression during viral (PI-3) infection [22]. The different levels of SBD-1 and surfactant protein A and D expression that occurs in M. haemolytica and PI-3 infections may influence the degree of clearance/persistence of each of these pathogens during pneumonia and may also influence susceptibility/resistance to secondary pathogens.…”

Section: Discussioncontrasting

confidence: 99%

“…SBD-1 and IL-8 gene expression were determined by two-step fluorogenic real time RT-PCR using primers and probe sequences (Table 2) listed, and according procedures previously described in our laboratory [15,22]. Briefly, real time RT-PCR was carried out according to manufacturer's specifications using the GeneAmp 5700 Sequence Detection System (Applied Biosystems).…”

Section: Real Time Rt-pcrmentioning

confidence: 99%

“…Captured cells were lysed and total RNA was isolated with a commercial kit (PicoPure RNA isolation kit; Arcturus, Inc.). SBD-1 and SBD-2 mRNA levels were determined by real time RT-PCR as described previously [15,22] and above. In order to determine if SBD-1 expression is present in leukocytes that infiltrate into the lung during pneumonia, SBD-1 and IL-8 levels were determined in buffy coat cells isolated from normal lambs.…”

Section: Real Time Rt-pcrmentioning

confidence: 99%

“…Total RNA was isolated from floating and trypsin-EDTA harvested cells using Trizol according to manufacturer's recommendations except our modification of using a 16,100g spin for the final RNA pellet collection (instead of 7500g). SBD-1 and IL-8 mRNA levels were determined by real time RT-PCR as described previously [15,22] and outlined above.…”

Section: Real Time Rt-pcrmentioning

confidence: 99%

“…SBD-1 is developmentally regulated in late gestation through the neonatal period with maximal expression in trachea, lung and rumen reached shortly after birth [19 -21]. Although SBD-1 appears to lack NF-kappa B responsive elements, [21][22][23][24][25][26][27] www.elsevier.com/locate/micpath SBD-1 expression is increased during infection with ovine parainfluenza virus type 3 (PI-3) [22]. SBD-2 is also developmentally regulated and expressed primarily in the alimentary tract [20,23]; however, only low levels of SBD-2 mRNA are present in lung relative to SBD-1 [20,23].…”

Section: Introductionmentioning

confidence: 99%

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Differential expression of sheep beta-defensin-1 and -2 and interleukin 8 during acute Mannheimia haemolytica pneumonia

Ackermann¹,

Gallup²,

Zabner³

et al. 2004

Microbial Pathogenesis

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Beta-defensins are antimicrobial peptides produced by several cell types, including respiratory epithelia and leukocytes. Expression of some beta-defensins is increased by bacterial-induced inflammatory responses whereas expression of other beta-defensins is constitutive. Two beta-defensins are expressed in lungs of sheep (sheep beta-defensin-1 and -2; SBD-1/-2) and expression of SBD-1 is increased during parainfluenza virus type 3 (PI-3) infection. The effect of Mannheimia haemolytica, a Gram-negative bacteria known to induce expression of bovine beta-defensins and NF-kappa B in lung, has not been determined for SBD-1/-2. In this study, different concentrations of M. haemolytica were inoculated into pulmonary bronchi of lambs. SBD-1 and SBD-2 mRNA levels detected by real time reverse transcriptase polymerase chain reaction in lung homogenates did not increase. In fact, SBD-1 mRNA levels were significantly decreased with the highest administered inoculum concentration (10 9 ). In contrast, mRNA levels of interleukin-8 (IL-8) were significantly increased over controls and progressively increased with M. haemolytica concentrations. Coinoculation of M. haemolytica with xylitol, an osmotic agent, did not alter mRNA levels of SBD-1, SBD-2 or IL-8. SBD-1 mRNA expression was detected in lung epithelia, but not in leukocytes. This study suggests that SDB-1 expression occurs in epithelia and decreases during severe bacterial pneumonia, which is in contrast to the increase that occurs with PI-3 infection. RightsWorks produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted. AuthorsMark R. Ackermann, Jack M. Abstract Beta-defensins are antimicrobial peptides produced by several cell types, including respiratory epithelia and leukocytes. Expression of some beta-defensins is increased by bacterial-induced inflammatory responses whereas expression of other beta-defensins is constitutive. Two beta-defensins are expressed in lungs of sheep (sheep beta-defensin-1 and -2; SBD-1/-2) and expression of SBD-1 is increased during parainfluenza virus type 3 (PI-3) infection. The effect of Mannheimia haemolytica, a Gram-negative bacteria known to induce expression of bovine beta-defensins and NF-kappa B in lung, has not been determined for SBD-1/-2. In this study, different concentrations of M. haemolytica were inoculated into pulmonary bronchi of lambs. SBD-1 and SBD-2 mRNA levels detected by real time reverse transcriptase polymerase chain reaction in lung homogenates did not increase. In fact, SBD-1 mRNA levels were significantly decreased with the highest administered inoculum concentration (10 9 ). In contrast, mRNA levels of interleukin-8 (IL-8) were significantly increased over controls and progressively increased with M. haemolytica concentrations. Co-inoculation of M. haemolytica with xylitol, an osmotic agent, did not alter mRNA levels of SBD-1, SBD-2 or IL-8. SBD-1 mRNA expression was detected in lung epithelia, b...

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Section: Discussioncontrasting

confidence: 99%

Section: Real Time Rt-pcrmentioning

confidence: 99%

Section: Real Time Rt-pcrmentioning

confidence: 99%

Section: Real Time Rt-pcrmentioning

confidence: 99%