EURURO Vol. 71 No. 4

with cavernous tissues cultured with autologous PBMCs

( Fig. 2 a

) (

= 0.0095). Following premaximal contraction

with PE, EFS-mediated relaxation was also impaired in

rejecting tissues comparedwith nonrejecting tissues

( Fig. 2

(

= 0.0148). Given the relatively short exposure time (48 h),

these data suggest that cavernous tissue physiological

function is sensitive to this model of rejection.

3.2.

Prevention of and characterization of rejection in the ex

vivo mixed lymphocyte reaction

Given the effects of rejection on corporal tissue function, we

next investigated whether penile tissue rejection could be

prevented by CsA, a well-known calcineurin inhibitor

commonly used for immunosuppression. Ex vivo MLR

was prepared as above with an additional group comprising

cavernous tissues cultured with allogenic PBMCs treated

with 1

M CsA

( Fig. 3 a

), a dose previously shown to prevent

PBMC activation in MLR rejection

[10] .

A human quantita-

tive PCR gene expression array assessing 84 transplant-

associated genes was used to further characterize the

rejection process in the ex vivo MLR and to determine

whether CsA treatment can prevent rejection

( Fig. 3

b and

3c). Array analysis normalized to autotransplant tissues

demonstrated global increased transplant-associated gene

expression in PBMCs exposed to allogenic cavernous tissue

compared with PBMCs exposed to autologous cavernous

tissue, which was attenuated by CsA immunosuppression

( Fig. 3

b). Pathway analysis demonstrated increased

activity in innate immunity, adaptive immunity, leukocyte

[(Fig._3)TD$FIG]

Fig. 3 – An additional experimental group (a) composed of an allotransplant (Allo) mixed lymphocyte reaction (MLR) treated with 1

M cyclosporine A

(CsA) was added to investigate the effects of immunosuppression treatment on penile rejection and function. (b) Heat map of an 84-gene polymerase

chain reaction human transplant array of peripheral blood mononuclear cells (PBMCs) removed from cavernous tissue MLR after 48 h of culture.

Expression was normalized to the autotransplant group. Allotransplant PBMCs demonstrated generalized increased expression of transplant-associated

genes. PBMCs treated with 1

M CsA demonstrated decreased transplant-associated gene expression compared with Allo. (c) Bar graphs of gene

expression of transplant-associated genes grouped by function demonstrated rejection occurring in the Allo group that was mitigated by CsA

treatment.

= 3 per group.

Allo = allotransplant; Auto = autotransplant; CsA = cyclosporine A; PBMC = peripheral blood mononuclear cell.

E U R O P E A N U R O L O G Y 7 1 ( 2 0 1 7 ) 5 8 4 – 5 9 3

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