Clostridium perfringens
-induced necrotic enteritis (
NE
) is an economically important disease of broiler chickens. The present study evaluated the effect of
C. perfringens
on the intestinal histomorphometry, enteric microbial colonization, and host immune responses using 3 experimental NE reproduction methods. The experimental groups consisted of 1) unchallenged Control diet (corn-soybean meal), 2) Control diet +
Eimera
inoculation at d 11 followed by
C. perfringens
challenge at d 15 (
ECp
), 3) Wheat-based diet +
C. perfringens
challenge (
WCp
), and 4) Wheat-based diet +
Eimeria
inoculation followed by
C. perfringens
challenge (
WECp
). The results showed that chickens receiving ECp and WECp had reduced (
P
< 0.05) bird performance coupled with enteric gross lesions and epithelial damage at d 17 and 24 of age compared to unchallenged control birds. These ECp and WECp administered birds also had increased (
P
< 0.05) ileal colonization by clostridia and
E. coli
at d 17 and 24, while the resident
Lactobacillus
counts were reduced (
P
< 0.05) at d 24 of age. Furthermore, at d 24, jejunal transcription of IL-6, IL-10, annexin-A1 and IL-2 genes was upregulated (
P
< 0.05) in the ECp group, whereas the transcription of TNF receptor associated factor (TRAF)-3 gene was increased (
P
< 0.05) in WECp treated birds when compared to unchallenged control group. Additionally, stimulation of chicken splenocytes and cecal tonsilocytes with virulent
C. perfringens
bacilli or their secretory proteins resulted in a higher (
P
< 0.05) frequency of T cells and their upregulation of MHC-II molecule, as determined by flow cytometry. These findings suggest that
C. perfringens
, while inducing epithelial damage and changes in microbiota, can also trigger host immune responses. Furthermore, NE reproduction methods using coccidia with or without the wheat-based dietary predisposition seem to facilitate an optimal NE reproduction in broiler chickens and thus, may provide better avenues for future
C. perfringens
research.