Early indicators of exposure to biological threat agents using host gene profiles in peripheral blood mononuclear cells

Das, Rina; Hammamieh, Rasha; Neill, Roger; Ludwig, George V.; Eker, Steven; Lincoln, Patrick; Ramamoorthy, Preveen; Dhokalia, Apsara; Mani, Sachin; Mendis, Chanaka; Cummings, Christiano; Kearney, Brian David; Royaee, Atabak R.; Huang, Xiaofang; Paranavitana, Chrysanthi; Smith, Leonard A.; Peel, Sheila A.; Kanesa-thasan, Niranjan; Hoover, David M.; Lindler, Luther E.; Yang, David C.H.; Henchal, Erik A.; Jett, Marti

doi:10.1186/1471-2334-8-104

Cited by 9 publications

(7 citation statements)

References 39 publications

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“…Human PBMC exposed to virus is a model for the viremic stage of infection in vivo when much of the transcriptome response is due to non-infectious signal transduction by viral particles [31] [24]. PBMC were used here and in other studies [32] [33] [34] rather than cell lines [35] [36] to more closely simulate the situation in vivo in which circulating blood cells are exposed to virus-infected sites. Here we have shown that in vitro PBMC exposure to arenaviruses with different pathogenic potential, LASV and ML29, resulted in differential expression of ISG, apoptotic, NF-kB, and coagulation pathway genes.…”

Section: Introductionmentioning

confidence: 99%

Transcriptome Analysis of Human Peripheral Blood Mononuclear Cells Exposed to Lassa Virus and to the Attenuated Mopeia/Lassa Reassortant 29 (ML29), a Vaccine Candidate

et al. 2013

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Lassa virus (LASV) is the causative agent of Lassa Fever and is responsible for several hundred thousand infections and thousands of deaths annually in West Africa. LASV and the non-pathogenic Mopeia virus (MOPV) are both rodent-borne African arenaviruses. A live attenuated reassortant of MOPV and LASV, designated ML29, protects rodents and primates from LASV challenge and appears to be more attenuated than MOPV. To gain better insight into LASV-induced pathology and mechanism of attenuation we performed gene expression profiling in human peripheral blood mononuclear cells (PBMC) exposed to LASV and the vaccine candidate ML29. PBMC from healthy human subjects were exposed to either LASV or ML29. Although most PBMC are non-permissive for virus replication, they remain susceptible to signal transduction by virus particles. Total RNA was extracted and global gene expression was evaluated during the first 24 hours using high-density microarrays. Results were validated using RT-PCR, flow cytometry and ELISA. LASV and ML29 elicited differential expression of interferon-stimulated genes (ISG), as well as genes involved in apoptosis, NF-kB signaling and the coagulation pathways. These genes could eventually serve as biomarkers to predict disease outcomes. The remarkable differential expression of thrombomodulin, a key regulator of inflammation and coagulation, suggests its involvement with vascular abnormalities and mortality in Lassa fever disease.

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Section: Introductionmentioning

confidence: 99%

Transcriptome Analysis of Human Peripheral Blood Mononuclear Cells Exposed to Lassa Virus and to the Attenuated Mopeia/Lassa Reassortant 29 (ML29), a Vaccine Candidate

et al. 2013

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“…The Trizol isolated RNA was used in cDNA microarrays analysis. 21 For oligonucleotide microarrays, total RNA was isolated using PAXgene tubes following the manufacturer's protocol. Isolated RNA samples were stored at –80 °C until they were used for microarray and real-time PCR analyses.…”

Section: Methodsmentioning

confidence: 99%

Transcriptome characterization of immune suppression from battlefield-like stress

et al. 2012

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Transcriptome alterations of leukocytes from soldiers who underwent 8 weeks of Army Ranger training (RASP, Ranger Assessment and Selection Program) were analyzed to evaluate impacts of battlefield-like stress on the immune response. About 1400 transcripts were differentially expressed between pre- and post-RASP leukocytes. Upon functional analysis, immune response was the most enriched biological process, and most of the transcripts associated with the immune response were downregulated. Microbial pattern recognition, chemotaxis, antigen presentation and T-cell activation were among the most downregulated immune processes. Transcription factors predicted to be stress-inhibited (IRF7, RELA, NFκB1, CREB1, IRF1 and HMGB) regulated genes involved in inflammation, maturation of dendritic cells and glucocorticoid receptor signaling. Many altered transcripts were predicted to be targets of stress-regulated microRNAs. Post-RASP leukocytes exposed ex vivo to Staphylococcal enterotoxin B showed a markedly impaired immune response to this superantigen compared with pre-RASP leukocytes, consistent with the suppression of the immune response revealed by transcriptome analyses. Our results suggest that suppression of antigen presentation and lymphocyte activation pathways, in the setting of normal blood cell counts, most likely contribute to the poor vaccine response, impaired wound healing and infection susceptibility associated with chronic intense stress.

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“…A laser detection system was used (GenePix 4000b, Axon Instruments, CA) to scan the finished slides. The intensity of the scanned images was digitalized through Genepix 4.0 software (Axon Inc., CA) [26].…”

Section: Materials and Methodologymentioning

confidence: 99%

“…Experiment was performed using 18S rRNA as the reference for quantitation as described in the Materials and Methods. The primer set was designed specifically for individual pathogen or toxin as described elsewhere(26).…”

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confidence: 99%

Identification of Early Response Genes in Human Peripheral Leukocytes Infected with Orientia tsutsugamushi: The Emergent of a Unique Gene Expression Profile for Diagnosis of O. tsutsugamush Infection~!2010-03-24~!2010-06-08~!2010-08-02~!

Chao¹,

Li²,

Hammamieh³

et al. 2010

TOIDJ

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Scrub typhus, caused by infection with Orientia tsutsugamushi, is one of the most common rickettsial diseases in the Asia-Pacific region. The disease can cause up to 35% mortality if left untreated. In order to get a better understanding of the host responses to O. tsutsugamushi infection, freshly isolated peripheral blood mononuclear cells (PBMC) were infected with O. tsutsugamushi. The infected cells were collected at 1 h, 4 h, 8 h or 18 h post infection. The gene expression profiles were monitored by cDNA microarray. Among the 7,489 genes, 658 genes were up or down regulated by 2-fold upon infection. ANOVA t-test revealed 432 genes with statistically significant fold change (p < 0.05). Semi-quantitative PCR using specific primers that flank the mRNA splicing sites of these 658 genes was carried out to verify the microarray results. More than 9000 semi-quantitative PCR reactions were performed using glyceraldehyde 3phosphate dehydrogenase (GAPDH) as the reference gene. Of these reactions, 22 genes were confirmed to exhibit up or down regulation. Quantitative PCR using 18S rRNA as the reference further confirmed two genes (NM_001547 and NM_006187) that exhibited significant increase of expression. Analysis of these 22 genes and the specific increase of both NM_001547 and NM_006187 suggest that the 22 genes are unique to O. tsutsugamushi infection and have the potential use for differentiating infections caused by virus, bacteria, parasites and other pathogens.

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Early indicators of exposure to biological threat agents using host gene profiles in peripheral blood mononuclear cells

Cited by 9 publications

References 39 publications

Transcriptome Analysis of Human Peripheral Blood Mononuclear Cells Exposed to Lassa Virus and to the Attenuated Mopeia/Lassa Reassortant 29 (ML29), a Vaccine Candidate

Transcriptome Analysis of Human Peripheral Blood Mononuclear Cells Exposed to Lassa Virus and to the Attenuated Mopeia/Lassa Reassortant 29 (ML29), a Vaccine Candidate

Transcriptome characterization of immune suppression from battlefield-like stress

Identification of Early Response Genes in Human Peripheral Leukocytes Infected with Orientia tsutsugamushi: The Emergent of a Unique Gene Expression Profile for Diagnosis of O. tsutsugamush Infection~!2010-03-24~!2010-06-08~!2010-08-02~!

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