The 'human challenge' study will see participants receive either the vaccine or a placebo.
Two to four months later, they will be deliberately exposed to the bacteria that causes whooping cough to find out how effective the proposed new vaccine is at stopping infection.
Participants, who will each be paid £3,775 to take part, will then be sent to a hotel for 16 nights.
It is not clear what hotel volunteers will be put up in but volunteers will have access to multiple rooms, a toilet, shower and recreational area. All meals, drinks and snacks will be provided.
They will also be allowed outside for two hours per day.
Before leaving the hotel two-and-a-half weeks later, the volunteers will be given a course of antibiotics to clear the bacteria in their nose.
Whooping cough causes coughing bouts that last for minutes at a time. It usually brings up thick mucus and may trigger vomiting.
Between coughs, sufferers may gasp for breath which causes a 'whoop' sound.
Anyone can be become infected. But adults usually have none or mild symptoms, while babies can suffer life-threatening dehydration, breathing difficulties and pneumonia.
A vaccine that protects against whooping cough is currently offered to all newborns in the UK.
However, it does not offer lifelong protection, cannot stop upper airway infections and does not prevent people from spreading it to others.
The current vaccine was introduced two decades ago to replace an older version that triggered side effects.
However, the jab used now is less effective. The UK has seen cases rise four-fold from 6,216 in the six years to 2011 to 25,891 in the six years to 2018.
And the infection is estimated to trigger 160,000 deaths worldwide per year.
Scientists hope the new vaccine will provide better protection and bring down infections.
Earlier studies found the nasal spray infected volunteers' nose and throat without causing any symptoms and produced an immune response.
Now, the human trial study will test the how well it can protect against the virus. After being exposed to the vaccine or placebo, volunteers will be infected in a research clinic.
Professor Robert Read, director of the National Institute for Health and Care Research Southampton Biomedical Research Centre, said: 'Despite the dramatic decline in whooping cough during the 20th century, there has been a sharp increase in cases in recent decades.
'There is an important need for new and more effective vaccines.
'We are inviting people in the Southampton area to become part of this journey to best protect people against this life-threatening bacteria.'