Gut health immunomodulatory and anti-inflammatory functions of gut enzyme digested high protein micro-nutrient dietary supplement-Enprocal

Table 3 Effect of Enprocal D and other digested milk product controls on modulation of cytokine secretion determined by ELISA.

Cells	Cytokines	Enprocal D	WPC	P2	Media +cells	Enzyme cocktail	LPS^a/PMA^b
THP-1 cells ^a	IL-1β	305 ± 10**	180 ± 8**	120 ± 5**	UD	700 ± 32^c	710 ± 35
	TNF-α	50 ± 2**	80 ± 2**	UD	UD	520 ± 27^c	530 ± 28
	IL-6	70 ± 5**	20 ± 2	10 ± 2	UD	114 ± 4^c	125 ± 4
	IL-8	1495 ± 15	1485 ± 12	1515 ± 16	UD	1500 ± 1^c	1505 ± 15
Jurkat cells ^b	IL-2	905 ± 12	925 ± 10	535 ± 14**	UD	UD^d	955 ± 15
	IFN-γ	105 ± 5	40 ± 5*	10 ± 2	UD	UD^d	81 ± 5
	IL-10	135 ± 10**	115 ± 10*	105 ± 6*	UD	UD^d	91 ± 6

UD = undetectable levels.
^aEnprocal digested and other digested milk product controls were added to Caco-2 cells in apical (upper) wells and culture supernatants were collected from the THP-1 (LPS stimulated) cells grown in basolateral (bottom) chamber after 8 hours (TNF-α) and 24 hours (IL-1β, IL-6 and IL-8) incubation.
^bEnprocal digested and other digested milk product controls were added to Caco-2 cells in apical wells and culture supernatants were collected from the Jurkat cells grown in basolateral chamber. Supernatant from Jurkat cells stimulated with PMA was used as positive control for the measurement of IL-2, INF-γ and IL-10 cytokines.
^cControl THP-1 cells with enzyme cocktail and with LPS stimulation.
^dControl Jurkat cells with enzyme cocktail and without PMA stimulation.
Concentration of cytokines in the culture medium was determined by ELISA as described in the Methods section. Cytokine concentrations in negative control (untreated cells) and growth media only were below the detection limits. All samples were assayed in triplicate and three independent experiments were performed. The mean for all experiment was calculated and presented as a mean ± SD values.
** Indicates a highly significant P < 0.001 value from the control with positive control (LPS/PMA) only. *Indicates a significant P < 0.05 value from the control with positive control (LPS/PMA) only.

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