Morphological and phenotypic characterization of immature MoDCs.

<p>(A) Light microscopy (20X) of freshly isolated monocyte. (B) Light microscopy (20X) of 4^th day culture of presumably immature MoDCs showing clusters of veiled cells with pseudopodia. (C) Freshly isolated monocytes and 5 day culture of MoDCs were analysed for phenotypic changes by evaluating mRNA expression levels of various TLRs and costimulatory genes by qRT-PCR assay. Results are expressed as fold change (log10) in mRNA transcription of monocytes and MoDCs. Data presented are mean ± standard deviation of values obtained from three independent experiments involving two cattle of Hallikar breed. *<i>p</i>< 0.05.**<i>p</i>< 0.01.</p

Kinetics of cytokine mRNA transcription from MoDCs treated with either BGs or LPS.

<p>RNA was extracted and gene transcription was quantified by qRT-PCR at 0, 6 and 12 h. Results are expressed as fold induction (log10) of cytokine mRNA transcription by BGs or LPS stimulated cells compared to the media treated (Med) cells. GAPDH was used as an internal control and mRNA levels at 0 h was used as calibrator. Histograms represent mean cytokine levels and bars represent standard deviation of three independent experiments. *p < 0.05, **p<0.01, ns = non-significant.</p

Characterization of <i>E</i>. <i>coli</i> ghosts by electron microscopy.

<p>(A) Field emission scanning electron micrograph (FE-SEM) of protein E lysed <i>E</i>. <i>coli</i> ghosts showing transmembrane tunnels, indicated by arrow heads. (B) FESEM of intact <i>E</i>. <i>coli</i> before lysis. (C) Transmission electron micrograph (TEM) of <i>E</i>. <i>coli</i> ghosts with a loss of cytoplasmic contents but intact cellular morphology. (D) TEM of intact <i>E</i>. <i>coli</i> prior to gene <i>E</i> induction.</p

Antigen presenting capacity of MoDCs determined by mixed lymphocyte reaction.

<p>(A) Bovine MoDCs (1.5 x 10⁴) were cultured with allogenic T cells (1.1 x 10⁵) and treated either with ConA, BGs, LPS or media (Med). The T cells alone stimulated either with LPS or media were kept as controls. (B) MoDCs were cultured with allogenic T cells (1.1 x 10⁵) of a different breed in a graded manner. The cultures were maintained for three days and subsequently, T cell proliferation was measured by the uptake of MTT dye and expressed as stimulation index. Data presented are mean ± standard deviation of values for MoDCs and allogenic T cells isolated from three animals. *p<0.05.**p<0.01.</p

Morphological and phenotypical features of mature bovine MoDCs.

<p>(A) Light microscopy (20X) of MoDCs after stimulation with BGs demonstrated large dendrite like pseudopodia (B) qRT-PCR analysis of MoDCs after stimulation with either BGs or LPS or treated with media (Med). Results are expressed as fold induction (log10) in mRNA transcription after BGs or LPS stimulation compared to the media treated (non-stimulated) cells. Gene expressions were normalized to GAPDH and mRNA levels at 0 h were used as calibrator. Data presented are mean ± standard deviation of three independent experiments involving two cattle of Hallikar breed. **p < 0.01, ***p<0.001, ns = non-significant.</p