Louis Dupree was born in Greenville, North Carolina in 1925. He attended Greenville High School until about 1943, enlisting in the armed forces before earning his diploma. He served in World War II, first as a merchant seaman, then as an officer in the 11th Airborne Division in the Philippines campaign and occupation of Japan. Dupree received the Mariner's Medal, Merchant Marine Combat Bar, Combat Infantry Badge, Purple Heart, and Bronze Star.

During 1947 to 1955, Dupree earned his bachelor's, master's, and doctorate degrees in Anthropology from Harvard. He began research in Central and South Asia in 1949. In the United States, Dupree taught and conducted research at Air University, Maxwell Air Force Base, Alabama, from 1953 to 1957. In 1957, he accepted a teaching appointment in the Anthropology Department at Pennsylvania State University. Dupree taught as a Visiting Professor at Kabul University (Afghanistan), Princeton University, and the United States Military Academy at West Point.

From 1959 to 1983, Dupree was a representative of the American Universities Field Staff (AUFS) in Afghanistan and Pakistan. He spent two years researching and writing under the AUFS program, returning to the United States every third year to lecture at the twelve universities sponsoring AUFS. Dupree was also director of several archaeological surveys and excavations in Afghanistan from 1959 to1983, sponsored by American institutions such as the American Museum of Natural History, the National Science Foundation, the American Philosophical Society, and the National Endowment for the Humanities. His excavation on upper Paleolithic sites at Aq Kupruk in northern Afghanistan won international acclaim.

In 1966, Dupree married Nancy Hatch in Afghanistan, whom he had met while she was writing a guidebook of the Bamiyan area of Afghanistan. The Duprees continued to intermittently live and work in Afghanistan.

While on AUFS business in Kabul, Afghanistan just prior to the 1978 Soviet Invasion, Dupree was imprisoned by Afghans and their Soviet advisors. He was interrogated at length about United States intelligence operations and his interactions with Afghans. He was ultimately released through the intervention of Afghan friends. The account of his imprisonment appears as a six-part series, in American University Field Staff Reports (1980).

As other nations became interested in the plight of Afghanistan, the Duprees were called upon to advise the parliaments or foreign ministries of Germany, France, Sweden, Norway, England, Austria, Pakistan, and to the United Nations High Commission on Refugees. Dupree frequently corresponded with Pakistan's president, General Mohammed Zia Ul-Haq. In the United States, Dupree acted as a consultant to the State Department, United States Agency for International Development (U.S. AID), the Peace Corps, Esso Pakistan Fertilizer Company, United Nations Development Programs, UNESCO, Helsinki Watch, Amnesty International, the World Bank, and other organizations.

In 1985, after teaching for one year at West Point Military Academy and another year at Princeton University, he joined the Program in Islamic and Arabian Development Studies at Duke University as a Senior Research Associate. He held concurrent appointments at Duke University as Visiting Professor of Anthropology, Political Science, and Public Policy Studies. He also taught at the University of North Carolina at Chapel Hill in the departments of Anthropology and Political Science.

The Islamic and Arabian Development Studies Program operated at Duke from 1977 to 1989 under the direction of Professor Ralph Braibanti. It funded the acquisition of library materials that supported its mission, facilitating the accession to the Duke libraries of two major Middle Eastern collections: the Joseph J. Malone Collection on Arabian Affairs and the Louis and Nancy Hatch Dupree Collection on Islamic Inner Asia. The Dupree Collection consists of about 5,000 items, 200 of which are located in the David M. Rubenstein Rare Book & Manuscript Library. Reflecting the careers and interests of their donors, the materials in the collection focus on the anthropology, art, archaeology, folklore, linguistics, and history of Afghanistan. There are also significant holdings on Islam, the Central Asian Republics, and South Asia.

A social activist, Dupree was Honorary Director and one of the founders of the Afghan Relief Committee. The beneficiaries of the Afghan Relief Committee were primarily Doctors Without Borders, Freedom Medical of Washington, D.C., Aide Medicale International, and Sainte Sud of Marseilles. Dupree was involved with the Afghanistan Action Committee at Duke University and corresponded with members of many other humanitarian assistance organizations.

Dupree and Nancy Hatch Dupree spent years researching and living in Afghanistan and planned to retire there. Together they researched and planned to write a book on Afghan refugees in Pakistan. By 1987, their collaborative work earned them a joint award, the International Rescue Committee's Bronze Medal for Service to Afghanistan Refugees. In 1988, the Duprees returned to Pakistan as Joint Fulbright Senior Scholars. Louis Dupree authored many books and over 200 articles. His articles appeared in various publications such as , the , the , the , the , , and the .

Some of the books authored by Dupree include: (1973), (1970), (1970), (1963), and (1956).

Louis Dupree died in 1989 at the age of 63. Nancy Hatch Dupree continued her work with Afghan refugees.