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Three children and a 29-year-old woman from the same family died after a minivan collided with a truck.
The accident happened on the A40 near Oxford Monday night, Thames Valley police said.
Officers were called in at 9.50pm to receive reports of the collision between a silver Subaru and an HGV near the railway bridge west of Oxford.
The woman and children, two girls aged four and eight and a boy aged six, were pronounced dead at the scene.
Two other Subaru passengers, a 30-year-old man and an 18-month-old girl, were taken to John Radcliffe Hospital in Oxford, where they are in critical condition.
The driver of the HGV, a 56-year-old man, suffered minor injuries.
The family was from Chinnor in Oxfordshire.
Sergeant Dominic Mahon, Senior Investigation Officer, Bicester-based Major Collision Investigation Unit, said the crash was an “absolute tragedy.”
He said: “We are in the early stages of our investigation into this incredibly tragic incident that has resulted in the deaths of four people from the same family.
“The closest family members have been informed and are supported by specially trained family liaison officers.
“The thoughts of all of us at Thames Valley Police, along with our partners who have assisted on the scene or at the hospital, are with families at this incredibly difficult time.”
He added: “We will leave no stone unturned to determine what has caused this tragedy.”
Officers remain at the scene and the A40 will be closed in both directions for a “considerable” period of time while the investigation continues.
Anyone in the area at the time of the collision and witnessed what happened should contact 101 or make a report online quoting reference number 43200321914
