England and Wales fans appear to have been caught up in violent clashes with Russian fans in video footage posted on social media.
Video posted on Twitter showed two sets of fans facing off outside a bar, with chairs strewn across the pavement on the Place de la Gare, in central Lille.
A group of people, some wearing football shirts, could be heard chanting "We're England and Wales, f--- off Russia" as police and security staff tried to calm the situation.
The footage shows a man in a suit shouting "allez," French for "go," to hooded and masked fans posturing towards the England and Wales supporters outside Le Palais de la Biere bar next to the city's Hotel Continental.
He attempts to set chairs and tables right again before police officers move in between the two groups of fans.
Members of the public could also be seen gathering and taking photos of the melee.
One England fan wearing a Rooney shirt was seen being led away in handcuffs by the police. The fans were heard once again singing songs about the IRA and German bombers.
A group of five fans wearing Russian flags and shirts attempted to walk over to England fans outside the Trois Brasseurs pub in Lille.
They were removed by police to the doors of the nearby Lille Flanders railway station where officers were seen checking their identities.
England fans were seen stamping on a Russian flag.
The fresh trouble comes after UEFA handed Russia a suspended disqualification from the tournament following the crowd disturbances at the game against the Three Lions in Marseille on Saturday.
The country was warned that any further disturbances from the nation's supporters would see them kicked out of the competition.
The Football Association (FA) were warned previously that further disorder could lead to the England team also being expelled from the tournament, even though the FA was not formally charged by the European football governing body.
A huge security operation surrounding England's next Euro 2016 football match has been bolstered, with extra British police being deployed to France ahead of the next round of Group B matches.
Authorities in both England and Russia have appealed for supporters to behave in the wake of disorder in Marseille.
French police arrested 43 Russian fans suspected of being involved in trouble at the weekend after stopping a bus in the Alpes-Maritimes region in the south of the country on Tuesday, according to local media reports.
And UK police are also assisting the investigation into the Marseille melee by scouring CCTV, social media and press images to identify any fans who were involved, the National Police Chiefs' Council (NPCC) said.
English fans suspected of disorder could face court action when they return to the UK in the form of football banning orders, the NPCC spokeswoman warned.
NPCC lead for football policing, assistant chief constable Mark Roberts appealed for witnesses to send police any footage and said: "A number of images of those involved in disorder have been collated and work continues to identify those involved."
Concerns have been raised about the security of fans as England prepare to take on Wales in Lens on Thursday and Russia play Slovakia in Lille on Wednesday, with the northern French cities just 23 miles apart.
The security operation in Lens, which one local official said would be "in lockdown," will be boosted by additional British police officers and also by British Transport Police, who will extend patrols from Eurostar trains to local rail services.
Fabienne Buccio, prefect of the Pas-de-Calais region, said 1,200 police officers and 1,200 private security workers would be on duty on the day of the game.
A ban on off-licences selling alcohol will also be in force in both cities between Tuesday evening and Friday morning.
Answering an urgent question in the Commons on Tuesday, Theresa May said British police officers known as "spotters" who are trained to identify troublemakers have been sent to France.
She said that for "operational reasons" she would not reveal how many officers are being sent across the Channel.
An NPCC spokeswoman said the additional officers being deployed for the England game against Wales had "already been agreed with the French" prior to the tournament, rather than a "reaction to what's happened recently".
Some 50,000 English and Welsh supporters are due to descend on Lens, while many fans are also expected to be staying in Lille and Mrs May said the Foreign Office is advising England fans without tickets not to travel to either city.
Six British fans were jailed for between one and three months on Monday for their part in the Marseille violence, while on Tuesday 24-year-old Birmingham City supporter Daniel Warlow became the first to to be given a five-year football banning order by a UK court for his part in the disorder on Friday, June 10.