James M. J. Sanno was assigned to command in August, and the regiment saw its first combat action in 1902 while serving as part of the American force sent to quell the Philippine Insurrection on the island of Mindanao.

This unit was involved in the August 19, 1904 riot with Ohio National Guard and civil authorities that resulted in several deaths.

After seeing extensive action in the Pacific theater during World War II, especially on the island of Guadalcanal during the Battle of Mount Austen, the Galloping Horse, and the Sea Horse, it fought in the last days of the New Georgia Campaign on the right flank on the advance on Munda, Solomon Islands, later during the Battle of Luzon and the ensuing occupation of Japan, the 27th Infantry Regiment earned the nickname "Gentle Wolfhounds" for their support of the Holy Family Home orphanage.

The unit saw heavy action throughout the war, where they were considered the "fire brigade" for the 25th Infantry Division – in essence, making first combat contact with enemy forces.

The 2nd Battalion, 27th Infantry Regiment, was again activated on 31 August 1995, and this unit again carries its thirty battle streamers and twelve-unit citations on its colors.

Both battalions of the 27th Infantry have entered the 71st year of the regiment's relationship with the children and staff of the Holy Family Home Orphanage, Osaka, Japan.

Recognizing the needs of the children, and the then-limited capacity for self-help in postwar Japan, the regiment turned what was to have been a one-time occurrence into flow of supplies, food, building materials, and medical assistance from American soldiers and their families to the orphans.

Soldiers from 1st and 2nd Battalions return to Japan every Christmas,[9] and two children from the orphanage have visited Schofield Barracks annually since 1957.

[10] The relationship was recognized by Hollywood in 1956 when members of the regiment were profiled in the 1955 film, Three Stripes in the Sun, starring Aldo Ray.

The movie Three Stripes in the Sun[12] (1955) is based on the New Yorker magazine article "The Gentle Wolfhound"[13] by Ely Jacques Kahn, Jr.

The movie "Lord of War" has a scene where a fascist soldier depicts the 27th Infantry Regiment crest on his beret while talking to Yuri Orlov.